Compounds and methods for reducing snca expression

ABSTRACT

Provided are compounds, methods, and pharmaceutical compositions for reducing the amount or activity of SNCA mRNA in a cell or animal, and in certain instances reducing the amount of alpha-synuclein protein in a cell or animal Such compounds, methods, and pharmaceutical compositions are useful to ameliorate at least one symptom or hallmark of a neurodegenerative disease. Such symptoms and hallmarks include motor dysfunction, aggregation of alpha-synuclein, neurodegeneration, cognitive decline and dementia. Such neurodegenerative diseases include Parkinson&#39;s disease, dementia with Lewy bodies, diffuse Lewy body disease, pure autonomic failure, multiple system atrophy, neuronopathic Gaucher&#39;s disease and Alzheimer&#39;s disease.

SEQUENCE LISTING

The present application is being filed along with a Sequence Listing inelectronic format. The Sequence Listing is provided as a file entitledBIOL0289USC1SEQ_ST25.txt, created on Nov. 8, 2021, which is 712 KB insize. The information in the electronic format of the sequence listingis incorporated herein by reference in its entirety.

FIELD

Provided are compounds, methods, and pharmaceutical compositions forreducing the amount or activity of alpha-synuclein (SNCA) mRNA in a cellor animal, and in certain instances reducing the amount ofalpha-synuclein protein in a cell or animal Such compounds, methods, andpharmaceutical compositions are useful to ameliorate at least onesymptom or hallmark of a neurodegenerative disease. Such symptoms andhallmarks include motor dysfunction, aggregation of alpha-synuclein,neurodegeneration, cognitive decline and dementia. Suchneurodegenerative diseases include Parkinson's disease, dementia withLewy bodies, diffuse Lewy body disease, pure autonomic failure, multiplesystem atrophy, neuronopathic Gaucher's disease and Alzheimer's disease.

BACKGROUND

Alpha-synuclein is a small, highly charged 140-amino acid residueprotein, predominantly expressed in central nervous system (CNS)neurons, where it is localized at presynaptic terminals in closeproximity to synaptic vesicles (Iwai, et al., Neuron. 1995. 14:467-475). Alpha-synuclein is encoded by the SNCA gene. Alpha-synucleincan associate with lipid membranes by forming amphipathic α-helices, asshown in vitro (Davidson, et al., J. Biol. Chem. 1998. 273: 9443-9449).Although the function of alpha-synuclein is still poorly understood,several studies suggest that it is involved in modulating synaptictransmission, the density of synaptic vesicles, and neuronal plasticity(Cabin et al., J. Neurosci. 2002. 22: 8797-8807). It has also beensuggested that alpha-synuclein may have a chaperone function, asindicated by its effectiveness in preventing aggregation of proteins inin vitro assays (Souza et al., FEBS Lett. 2000. 474: 116-119). Moreover,in vivo assays demonstrate that alpha-synuclein chaperone activity isinstrumental in promoting the assembly of the SNARE-complex, which isessential for neurotransmitter release in the presynaptic terminals ofthe brain (Burre et al., Science. 329: 1663-1667). DecreasedSNARE-complex assembly is associated with neurological impairment, thus,indicating a link between presynaptic alpha-synuclein aggregates andneurodegeneration (Kramer and Schulz-Schaeffer, J. Neurosci. 2007. 27:1405-1410). Knockout mouse models of alpha-synuclein are not lethal, andbrain morphology is intact, suggesting that alpha-synuclein is notrequired for neuronal development and/or that compensatory pathways arepresent (Abeliovich et al., Neuron. 2000. 25: 239-252).

Misfolding, aggregation, and fibrillation of alpha-synuclein areimplicated as critical factors in several neurodegenerative diseases,including, Parkinson's disease, Lewy body variant of Alzheimer'sdisease, diffuse Lewy body disease, dementia with Lewy bodies, andmultiple system atrophy (Schulz-Schaeffer Acta Neuropathol. 2010. 120:131-143; Yoshida. Neuropathology. 2007. 27: 484-493). In each of thesecases, alpha-synuclein protein is misfolded and assembles in aggregatesin Lewy bodies and Lewy neurites (Uversky. J. Neurochem. 2007. 103:17-37). Several recent studies have shown that lipidic environments thatpromote alpha-synuclein folding also accelerate alpha-synucleinaggregation, suggesting that the lipid-associated conformation ofalpha-synuclein may be relevant to alpha-synuclein misfolding inneurodegenerative diseases (Conway et al., Science. 2001. 294: 6-9; Leeet al., J. Biol. Chem. 2002. 277: 671-678). Mutations at position 53,where alanine is changed to threonine, and at position 30, where alanineis changed to proline, have been shown to cause alpha-synuclein to be ina random coil state, so that aggregation is more likely to occur(Clayton and George, J. Neurosci. 1999. 58: 120-129).

Currently there is a lack of acceptable options for treatingneurodegenerative disease such as Parkinson's disease, dementia withLewy bodies, diffuse Lewy body disease, pure autonomic failure, multiplesystem atrophy, neuronopathic Gaucher's disease and Alzheimer's disease.It is therefore an object herein to provide compounds, methods, andpharmaceutical compositions for the treatment of such diseases.

SUMMARY OF THE INVENTION

Provided herein are compounds, methods and pharmaceutical compositionsfor reducing the amount or activity of SNCA mRNA, and in certainembodiments reducing the amount of alpha-synuclein protein in a cell oranimal. In certain embodiments, the animal has a neurodegenerativedisease. In certain embodiments, the animal has Parkinson's disease,dementia with Lewy bodies, diffuse Lewy body disease, pure autonomicfailure, multiple system atrophy, neuronopathic Gaucher's disease orAlzheimer's disease. In certain embodiments, compounds useful forreducing expression of SNCA mRNA are oligomeric compounds. In certainembodiments, compounds useful for reducing expression of SNCA mRNA aremodified oligonucleotides.

Also provided are methods useful for ameliorating at least one symptomor hallmark of a neurodegenerative disease. In certain embodiments, theneurodegenerative disease is Parkinson's disease, dementia with Lewybodies, diffuse Lewy body disease, pure autonomic failure, multiplesystem atrophy, neuronopathic Gaucher's disease and Alzheimer's disease.In certain embodiments, the symptom or hallmark includes motordysfunction, aggregation of alpha-synuclein, neurodegeneration,cognitive decline and dementia. In certain embodiments, amelioration ofthese symptoms results in improved motor function, reduction ofalpha-synuclein aggregates, reduced neurodegeneration and/or reduceddementia.

DETAILED DESCRIPTION OF THE INVENTION

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory onlyand are not restrictive. Herein, the use of the singular includes theplural unless specifically stated otherwise. As used herein, the use of“or” means “and/or” unless stated otherwise. Furthermore, the use of theterm “including” as well as other forms, such as “includes” and“included”, is not limiting. Also, terms such as “element” or“component” encompass both elements and components comprising one unitand elements and components that comprise more than one subunit, unlessspecifically stated otherwise.

The section headings used herein are for organizational purposes onlyand are not to be construed as limiting the subject matter described.All documents, or portions of documents, cited in this application,including, but not limited to, patents, patent applications, articles,books, and treatises, are hereby expressly incorporated-by-reference forthe portions of the document discussed herein, as well as in theirentirety.

Definitions

Unless specific definitions are provided, the nomenclature used inconnection with, and the procedures and techniques of, analyticalchemistry, synthetic organic chemistry, and medicinal and pharmaceuticalchemistry described herein are those well known and commonly used in theart. Where permitted, all patents, applications, published applicationsand other publications and other data referred to throughout in thedisclosure are incorporated by reference herein in their entirety.

Unless otherwise indicated, the following terms have the followingmeanings:

Definitions

As used herein, “2′-deoxynucleoside” means a nucleoside comprising a2′-H(H) deoxyribosy sugar moiety, as found in naturally occurringdeoxyribonucleic acids (DNA). In certain embodiments, a2′-deoxynucleoside may comprise a modified nucleobase or may comprise anRNA nucleobase (uracil).

As used herein, “2′-substituted nucleoside” means a nucleosidecomprising a 2′-substituted sugar moiety. As used herein,“2′-substituted” in reference to a sugar moiety means a sugar moietycomprising at least one 2′-substituent group other than H or OH.

As used herein, “5-methyl cytosine” means a cytosine modified with amethyl group attached to the 5 position. A 5-methyl cytosine is amodified nucleobase.

As used herein, “administering” means providing a pharmaceutical agentto an animal.

As used herein, “animal” means a human or non-human animal.

As used herein, “antisense activity” means any detectable and/ormeasurable change attributable to the hybridization of an antisensecompound to its target nucleic acid. In certain embodiments, antisenseactivity is a decrease in the amount or expression of a target nucleicacid or protein encoded by such target nucleic acid compared to targetnucleic acid levels or target protein levels in the absence of theantisense compound.

As used herein, “antisense compound” means an oligomeric compoundcapable of achieving at least one antisense activity.

As used herein, “ameliorate” in reference to a treatment meansimprovement in at least one symptom relative to the same symptom in theabsence of the treatment. In certain embodiments, amelioration is thereduction in the severity or frequency of a symptom or the delayed onsetor slowing of progression in the severity or frequency of a symptom. Incertain embodiments, the symptom or hallmark is motor dysfunction,aggregation of alpha-synuclein, neurodegeneration, cognitive declineand/or dementia. In certain embodiments, amelioration of these symptomsresults in improved motor function, reduction of alpha-synucleinaggregates, reduced neurodegeneration and/or reduced dementia.

As used herein, “bicyclic nucleoside” or “BNA” means a nucleosidecomprising a bicyclic sugar moiety.

As used herein, “bicyclic sugar” or “bicyclic sugar moiety” means amodified sugar moiety comprising two rings, wherein the second ring isformed via a bridge connecting two of the atoms in the first ringthereby forming a bicyclic structure. In certain embodiments, the firstring of the bicyclic sugar moiety is a furanosyl moiety. In certainembodiments, the bicyclic sugar moiety does not comprise a furanosylmoiety.

As used herein, “cleavable moiety” means a bond or group of atoms thatis cleaved under physiological conditions, for example, inside a cell,an animal, or a human.

As used herein, “complementary” in reference to an oligonucleotide meansthat at least 70% of the nucleobases of the oligonucleotide or one ormore regions thereof and the nucleobases of another nucleic acid or oneor more regions thereof are capable of hydrogen bonding with one anotherwhen the nucleobase sequence of the oligonucleotide and the othernucleic acid are aligned in opposing directions. Complementarynucleobases means nucleobases that are capable of forming hydrogen bondswith one another. Complementary nucleobase pairs include adenine (A) andthymine (T), adenine (A) and uracil (U), cytosine (C) and guanine (G),5-methyl cytosine (mC) and guanine (G). Complementary oligonucleotidesand/or nucleic acids need not have nucleobase complementarity at eachnucleoside. Rather, some mismatches are tolerated. As used herein,“fully complementary” or “100% complementary” in reference tooligonucleotides means that oligonucleotides are complementary toanother oligonucleotide or nucleic acid at each nucleoside of theoligonucleotide.

As used herein, “conjugate group” means a group of atoms that isdirectly or indirectly attached to an oligonucleotide. Conjugate groupsinclude a conjugate moiety and a conjugate linker that attaches theconjugate moiety to the oligonucleotide.

As used herein, “conjugate linker” means a group of atoms comprising atleast one bond that connects a conjugate moiety to an oligonucleotide.

As used herein, “conjugate moiety” means a group of atoms that isattached to an oligonucleotide via a conjugate linker.

As used herein, “contiguous” in the context of an oligonucleotide refersto nucleosides, nucleobases, sugar moieties, or internucleoside linkagesthat are immediately adjacent to each other. For example, “contiguousnucleobases” means nucleobases that are immediately adjacent to eachother in a sequence.

As used herein, “constrained ethyl” or “cEt” or “cEt modified sugar”means a β-D ribosyl bicyclic sugar moiety wherein the second ring of thebicyclic sugar is formed via a bridge connecting the 4′-carbon and the2′-carbon of the β-D ribosyl sugar moiety, wherein the bridge has theformula 4′-CH(CH₃)—O-2′, and wherein the methyl group of the bridge isin the S configuration.

As used herein, “cEt nucleoside” means a nucleoside comprising cEtmodified sugar.

As used herein, “chirally enriched population” means a plurality ofmolecules of identical molecular formula, wherein the number orpercentage of molecules within the population that contain a particularstereochemical configuration at a particular chiral center is greaterthan the number or percentage of molecules expected to contain the sameparticular stereochemical configuration at the same particular chiralcenter within the population if the particular chiral center werestereorandom. Chirally enriched populations of molecules having multiplechiral centers within each molecule may contain one or more stereorandomchiral centers. In certain embodiments, the molecules are modifiedoligonucleotides. In certain embodiments, the molecules are compoundscomprising modified oligonucleotides.

As used herein, “gapmer” means a modified oligonucleotide comprising aninternal region having a plurality of nucleosides that support RNase Hcleavage positioned between external regions having one or morenucleosides, wherein the nucleosides comprising the internal region arechemically distinct from the nucleoside or nucleosides comprising theexternal regions. The internal region may be referred to as the “gap”and the external regions may be referred to as the “wings.” Unlessotherwise indicated, “gapmer” refers to a sugar motif. Unless otherwiseindicated, the sugar moieties of the nucleosides of the gap of a gapmerare unmodified 2′-deoxyribosyl. Thus, the term “MOE gapmer” indicates agapmer having a sugar motif of 2′-MOE nucleosides in both wings and agap of 2′-deoxynucleosides. Unless otherwise indicated, a MOE gapmer maycomprise one or more modified internucleoside linkages and/or modifiednucleobases and such modifications do not necessarily follow the gapmerpattern of the sugar modifications.

As used herein, “hotspot region” is a range of nucleobases on a targetnucleic acid amenable to oligomeric compound-mediated reduction of theamount or activity of the target nucleic acid.

As used herein, “hybridization” means the pairing or annealing ofcomplementary oligonucleotides and/or nucleic acids. While not limitedto a particular mechanism, the most common mechanism of hybridizationinvolves hydrogen bonding, which may be Watson-Crick, Hoogsteen orreversed Hoogsteen hydrogen bonding, between complementary nucleobases.

As used herein, the term “internucleoside linkage” is the covalentlinkage between adjacent nucleosides in an oligonucleotide. As usedherein “modified internucleoside linkage” means any internucleosidelinkage other than a phosphodiester internucleoside linkage.“Phosphorothioate internucleoside linkage” is a modified internucleosidelinkage in which one of the non-bridging oxygen atoms of aphosphodiester internucleoside linkage is replaced with a sulfur atom.

As used herein, “linker-nucleoside” means a nucleoside that links,either directly or indirectly, an oligonucleotide to a conjugate moiety.Linker-nucleosides are located within the conjugate linker of anoligomeric compound. Linker-nucleosides are not considered part of theoligonucleotide portion of an oligomeric compound even if they arecontiguous with the oligonucleotide.

As used herein, “non-bicyclic modified sugar moiety” means a modifiedsugar moiety that comprises a modification, such as a substituent, thatdoes not form a bridge between two atoms of the sugar to form a secondring.

As used herein, “mismatch” or “non-complementary” means a nucleobase ofa first oligonucleotide that is not complementary with the correspondingnucleobase of a second oligonucleotide or target nucleic acid when thefirst and second oligonucleotide are aligned.

As used herein, “MOE” means methoxyethyl. “2′-MOE” or “2′-MOE modifiedsugar” means a 2′-OCH₂CH₂OCH₃ group in place of the 2′-OH group of aribosyl sugar moiety.

As used herein, “2′-MOE nucleoside” means a nucleoside comprising a2′-MOE modified sugar As used herein, “motif” means the pattern ofunmodified and/or modified sugar moieties, nucleobases, and/orinternucleoside linkages, in an oligonucleotide.

As used herein, “mRNA” means an RNA transcript that encodes a proteinand includes pre-mRNA and mature mRNA unless otherwise specified.

As used herein, “neurodegenerative disease” means a condition marked byprogressive loss of structure or function of neurons, including death ofneurons. In certain embodiments, the neurodegenerative disease isParkinson's disease, dementia with Lewy bodies, diffuse Lewy bodydisease, pure autonomic failure, multiple system atrophy, neuronopathicGaucher's disease and Alzheimer's disease.

As used herein, “nucleobase” means an unmodified nucleobase or amodified nucleobase. As used herein an “unmodified nucleobase” isadenine (A), thymine (T), cytosine (C), uracil (U), and guanine (G). Asused herein, a “modified nucleobase” is a group of atoms other thanunmodified A, T, C, U, or G capable of pairing with at least oneunmodified nucleobase. A “5-methyl cytosine” is a modified nucleobase. Auniversal base is a modified nucleobase that can pair with any one ofthe five unmodified nucleobases. As used herein, “nucleobase sequence”means the order of contiguous nucleobases in a nucleic acid oroligonucleotide independent of any sugar or internucleoside linkagemodification.

As used herein, “nucleoside” means a compound comprising a nucleobaseand a sugar moiety. The nucleobase and sugar moiety are each,independently, unmodified or modified. As used herein, “modifiednucleoside” means a nucleoside comprising a modified nucleobase and/or amodified sugar moiety. Modified nucleosides include abasic nucleosides,which lack a nucleobase. “Linked nucleosides” are nucleosides that areconnected in a contiguous sequence (i.e., no additional nucleosides arepresented between those that are linked).

As used herein, “oligomeric compound” means an oligonucleotide andoptionally one or more additional features, such as a conjugate group orterminal group. An oligomeric compound may be paired with a secondoligomeric compound that is complementary to the first oligomericcompound or may be unpaired. A “singled-stranded oligomeric compound” isan unpaired oligomeric compound. The term “oligomeric duplex” means aduplex formed by two oligomeric compounds having complementarynucleobase sequences. Each oligomeric compound of an oligomeric duplexmay be referred to as a “duplexed oligomeric compound.”

As used herein, “oligonucleotide” means a strand of linked nucleosidesconnected via internucleoside linkages, wherein each nucleoside andinternucleoside linkage may be modified or unmodified. Unless otherwiseindicated, oligonucleotides consist of 8-50 linked nucleosides. As usedherein, “modified oligonucleotide” means an oligonucleotide, wherein atleast one nucleoside or internucleoside linkage is modified. As usedherein, “unmodified oligonucleotide” means an oligonucleotide that doesnot comprise any nucleoside modifications or internucleosidemodifications.

As used herein, “pharmaceutically acceptable carrier or diluent” meansany substance suitable for use in administering to an animal Certainsuch carriers enable pharmaceutical compositions to be formulated as,for example, tablets, pills, dragees, capsules, liquids, gels, syrups,slurries, suspension and lozenges for the oral ingestion by a subject.In certain embodiments, a pharmaceutically acceptable carrier or diluentis sterile water, sterile saline, sterile buffer solution or sterileartificial cerebrospinal fluid.

As used herein “pharmaceutically acceptable salts” means physiologicallyand pharmaceutically acceptable salts of compounds. Pharmaceuticallyacceptable salts retain the desired biological activity of the parentcompound and do not impart undesired toxicological effects thereto.

As used herein “pharmaceutical composition” means a mixture ofsubstances suitable for administering to a subject. For example, apharmaceutical composition may comprise an oligomeric compound and asterile aqueous solution. In certain embodiments, a pharmaceuticalcomposition shows activity in free uptake assay in certain cell lines.

As used herein “prodrug” means a therapeutic agent in a form outside thebody that is converted to a different form within an animal or cellsthereof. Typically conversion of a prodrug within the animal isfacilitated by the action of an enzymes (e.g., endogenous or viralenzyme) or chemicals present in cells or tissues and/or by physiologicconditions.

As used herein, “reducing or inhibiting the amount or activity” refersto a reduction or blockade of the transcriptional expression or activityrelative to the transcriptional expression or activity in an untreatedor control sample and does not necessarily indicate a total eliminationof transcriptional expression or activity.

As used herein, “RNAi compound” means an antisense compound that acts,at least in part, through RISC or Ago2 to modulate a target nucleic acidand/or protein encoded by a target nucleic acid. RNAi compounds include,but are not limited to double-stranded siRNA, single-stranded RNA(ssRNA), and microRNA, including microRNA mimics. In certainembodiments, an RNAi compound modulates the amount, activity, and/orsplicing of a target nucleic acid. The term RNAi compound excludesantisense compounds that act through RNase H.

As used herein, “self-complementary” in reference to an oligonucleotidemeans an oligonucleotide that at least partially hybridizes to itself.

As used herein, “standard cell assay” means the assay described inExample 10 and reasonable variations thereof.

As used herein, “standard in vivo assay” means the experiment describedin Example 22 and reasonable variations thereof.

As used herein, “stereorandom chiral center” in the context of apopulation of molecules of identical molecular formula means a chiralcenter having a random stereochemical configuration. For example, in apopulation of molecules comprising a stereorandom chiral center, thenumber of molecules having the (S) configuration of the stereorandomchiral center may be but is not necessarily the same as the number ofmolecules having the (R) configuration of the stereorandom chiralcenter. The stereochemical configuration of a chiral center isconsidered random when it is the results of a synthetic method that isnot designed to control the stereochemical configuration. In certainembodiments, a stereorandom chiral center is a stereorandomphosphorothioate internucleoside linkage.

As used herein, “sugar moiety” means an unmodified sugar moiety or amodified sugar moiety. As used herein, “unmodified sugar moiety” means a2′-OH(H) ribosyl moiety, as found in RNA (an “unmodified RNA sugarmoiety”), or a 2′-H(H) deoxyribosyl moiety, as found in DNA (an“unmodified DNA sugar moiety”). Unmodified sugar moieties have onehydrogen at each of the 1′, 3′, and 4′ positions, an oxygen at the 3′position, and two hydrogens at the 5′ position. As used herein,“modified sugar moiety” or “modified sugar” means a modified furanosylsugar moiety or a sugar surrogate.

As used herein, “sugar surrogate” means a modified sugar moiety havingother than a furanosyl moiety that can link a nucleobase to anothergroup, such as an internucleoside linkage, conjugate group, or terminalgroup in an oligonucleotide. Modified nucleosides comprising sugarsurrogates can be incorporated into one or more positions within anoligonucleotide and such oligonucleotides are capable of hybridizing tocomplementary oligomeric compounds or target nucleic acids.

As used herein, “target nucleic acid” and “target RNA” mean a nucleicacid that an antisense compound is designed to affect.

As used herein, “target region” means a portion of a target nucleic acidto which an oligomeric compound is designed to hybridize.

As used herein, “terminal group” means a chemical group or group ofatoms that is covalently linked to a terminus of an oligonucleotide.

As used herein, “therapeutically effective amount” means an amount of apharmaceutical agent that provides a therapeutic benefit to an animal.For example, a therapeutically effective amount improves a symptom of adisease.

The present disclosure provides the following non-limiting numberedembodiments:

Embodiment 1. An oligomeric compound comprising a modifiedoligonucleotide consisting of 10-30 linked nucleosides and having anucleobase sequence comprising at least 12, 13, 14, 15, 16 or 17nucleobases of any of SEQ ID NOS: 2193, 1703, 28-1702, 1704-2192, and2194-2793.

Embodiment 2. An oligomeric compound comprising a modifiedoligonucleotide consisting of 10-30 linked nucleosides and having anucleobase sequence complementary to at least 8, at least 9, at least10, at least 11, at least 12, at least 13, at least 14, at least 15, atleast 16, at least 17, at least 18, at least 19, or at least 20contiguous nucleobases of:

an equal length portion of nucleobases 50915-50943 of SEQ ID NO: 2;

an equal length portion of nucleobases 19630-19656 of SEQ ID NO: 2;

an equal length portion of nucleobases 28451-28491 of SEQ ID NO: 2;

an equal length portion of nucleobases 48712-48760 of SEQ ID NO: 2;

an equal length portion of nucleobases 23279-23315 of SEQ ID NO: 2;

an equal length portion of nucleobases 20964-21018 of SEQ ID NO: 2;

an equal length portion of nucleobases 22454-22477 of SEQ ID NO: 2;

an equal length portion of nucleobases 72294-72321 of SEQ ID NO: 2;

an equal length portion of nucleobases 20549-20581 of SEQ ID NO: 2; oran equal length portion of nucleobases 27412-27432 of SEQ ID NO: 2.

Embodiment 3. The oligomeric compound of embodiment 1 or 2, wherein themodified oligonucleotide has a nucleobase sequence that is at least 80%,85%, 90%, 95%, or 100% complementary to any of the nucleobase sequencesof SEQ ID NO: 1-6, when measured across the entire nucleobase sequenceof the modified oligonucleotide.

Embodiment 4. The oligomeric compound of any of embodiments 1-3, whereinthe modified oligonucleotide comprises at least one modified nucleoside.

Embodiment 5. The oligomeric compound of embodiment 4, wherein themodified oligonucleotide comprises at least one modified nucleosidecomprising a modified sugar moiety.

Embodiment 6. The oligomeric compound of embodiment 5, wherein themodified oligonucleotide comprises at least one modified nucleosidecomprising a bicyclic sugar moiety.

Embodiment 7. The oligomeric compound of embodiment 6, wherein themodified oligonucleotide comprises at least one modified nucleosidecomprising a bicyclic sugar moiety having a 2′-4′ bridge, wherein the2′-4′ bridge is selected from —O—CH₂—; and —O—CH(CH₃)—.

Embodiment 8. The oligomeric compound of any of embodiments 4-7, whereinthe modified oligonucleotide comprises at least one modified nucleosidecomprising a non-bicyclic modified sugar moiety.

Embodiment 9. The oligomeric compound of embodiment 8, wherein themodified oligonucleotide comprises at least one modified nucleosidecomprising a non-bicyclic modified sugar moiety comprising a 2′-MOEmodified sugar or 2′-OMe modified sugar.

Embodiment 10. The oligomeric compound of any of embodiments 4-9,wherein the modified oligonucleotide comprises at least one modifiednucleoside comprising a sugar surrogate.

Embodiment 11. The oligomeric compound of embodiment 10, wherein themodified oligonucleotide comprises at least one modified nucleosidecomprising a sugar surrogate selected from morpholino and PNA.

Embodiment 12. The oligomeric compound of any of embodiments 1-11,wherein the modified oligonucleotide has a sugar motif comprising:

a 5′-region consisting of 1-5 linked 5′-region nucleosides;

a central region consisting of 6-10 linked central region nucleosides;and

a 3′-region consisting of 1-5 linked 3′-region nucleosides; wherein

each of the 5′-region nucleosides and each of the 3′-region nucleosidescomprises a modified sugar moiety and each of the central regionnucleosides comprises an unmodified 2′-deoxyribosyl sugar moiety.

Embodiment 13. The oligomeric compound of any of embodiments 1-12,wherein the modified oligonucleotide comprises at least one modifiedinternucleoside linkage.

Embodiment 14. The oligomeric compound of embodiment 13, wherein eachinternucleoside linkage of the modified oligonucleotide is a modifiedinternucleoside linkage.

Embodiment 15. The oligomeric compound of embodiment 13 or 14 wherein atleast one internucleoside linkage is a phosphorothioate internucleosidelinkage.

Embodiment 16. The oligomeric compound of embodiment 13 or 15 whereinthe modified oligonucleotide comprises at least one phosphodiesterinternucleoside linkage.

Embodiment 17. The oligomeric compound of any of embodiments 13, 15, or16, wherein each internucleoside linkage is either a phosphodiesterinternucleoside linkage or a phosphorothioate internucleoside linkage.

Embodiment 18. The oligomeric compound of any of embodiments 1-17,wherein the modified oligonucleotide comprises at least one modifiednucleobase.

Embodiment 19. The oligomeric compound of embodiment 18, wherein themodified nucleobase is a 5-methyl cytosine.

Embodiment 20. The oligomeric compound of any of embodiments 1-19,wherein the modified oligonucleotide consists of 12-30, 12-22, 12-20,14-20, 15-25, 16-20, 18-22 or 18-20 linked nucleosides.

Embodiment 21. The oligomeric compound of any of embodiments 1-20,wherein the modified oligonucleotide consists of 17 or 20 linkednucleosides.

Embodiment 22. The oligomeric compound of any of embodiments 1-21consisting of the modified oligonucleotide.

Embodiment 23. The oligomeric compound of any of embodiments 1-21comprising a conjugate group comprising a conjugate moiety and aconjugate linker.

Embodiment 24. The oligomeric compound of embodiment 23, wherein theconjugate group comprises a GalNAc cluster comprising 1-3 GalNAcligands.

Embodiment 25. The oligomeric compound of embodiment 23 or 24, whereinthe conjugate linker consists of a single bond.

Embodiment 26. The oligomeric compound of embodiment 24, wherein theconjugate linker is cleavable.

Embodiment 27. The oligomeric compound of embodiment 26, wherein theconjugate linker comprises 1-3 linker-nucleosides.

Embodiment 28. The oligomeric compound of any of embodiments 23-27,wherein the conjugate group is attached to the modified oligonucleotideat the 5′-end of the modified oligonucleotide.

Embodiment 29. The oligomeric compound of any of embodiments 23-27,wherein the conjugate group is attached to the modified oligonucleotideat the 3′-end of the modified oligonucleotide.

Embodiment 30. The oligomeric compound of any of embodiments 1-29comprising a terminal group.

Embodiment 31. The oligomeric compound of any of embodiments 1-30wherein the oligomeric compound is a singled-stranded oligomericcompound.

Embodiment 32. The oligomeric compound of any of embodiments 1-26 or28-30, wherein the oligomeric compound does not compriselinker-nucleosides.

Embodiment 33. An oligomeric duplex comprising an oligomeric compound ofany of embodiments 1-30 or 32.

Embodiment 34. An antisense compound comprising or consisting of anoligomeric compound of any of embodiments 1-32 or an oligomeric duplexof embodiment 33.

Embodiment 35. A pharmaceutical composition comprising an oligomericcompound of any of embodiments 1-32 or an oligomeric duplex ofembodiment 33 and a pharmaceutically acceptable carrier or diluent.

Embodiment 36. A modified oligonucleotide according to the followingformula:

or a salt thereof.Consistent with the definitions and disclosure herein, compound ofEmbodiment 36 may be made by deliberately controlling stereochemistry ofany, all or none of the linkages.

Embodiment 37. A modified oligonucleotide according to the followingformula:

or a salt thereof.Consistent with the definitions and disclosure herein, compound ofEmbodiment 37 may be made by deliberately controlling stereochemistry ofany, all or none of the linkages.

Embodiment 38. A modified oligonucleotide according to the followingformula:

or a salt thereof.Consistent with the definitions and disclosure herein, compound ofEmbodiment 38 may be made by deliberately controlly stereochemistry ofany, all or none of the linkages.

Embodiment 39. A modified oligonucleotide according to the followingformula:

or a salt thereof.Consistent with the definitions and disclosure herein, compound ofEmbodiment 39 may be made by deliberately controlling stereochemistry ofany, all or none of the linkages.

Embodiment 40. A modified oligonucleotide according to the followingformula:

or a salt thereof.Consistent with the definitions and disclosure herein, compound ofEmbodiment 40 may be made by deliberately controlling stereochemistry ofany, all or none of the linkages.

Embodiment 41. The modified oligonucleotide of any of embodiments 36-40,which is a sodium salt of the formula.

Embodiment 42. A chirally enriched population of the modifiedoligonucleotide of any of embodiments 36-40 wherein the population isenriched for modified oligonucleotides comprising at least oneparticular phosphorothioate internucleoside linkage having a particularstereochemical configuration.

Embodiment 43. The chirally enriched population of embodiment 42,wherein the population is enriched for modified oligonucleotidescomprising at least one particular phosphorothioate internucleosidelinkage having the (Sp) configuration.

Embodiment 44. The chirally enriched population of embodiment 42,wherein the population is enriched for modified oligonucleotidescomprising at least one particular phosphorothioate internucleosidelinkage having the (Rp) configuration.

Embodiment 45. The chirally enriched population of embodiment 42,wherein the population is enriched for modified oligonucleotides havinga particular, independently selected stereochemical configuration ateach phosphorothioate internucleoside linkage

Embodiment 46. The chirally enriched population of embodiment 45,wherein the population is enriched for modified oligonucleotides havingthe (Sp) configuration at each phosphorothioate internucleoside linkage.

Embodiment 47. The chirally enriched population of embodiment 45,wherein the population is enriched for modified oligonucleotides havingthe (Rp) configuration at each phosphorothioate internucleoside linkage.

Embodiment 48. The chirally enriched population of embodiment 45,wherein the population is enriched for modified oligonucleotides havingthe (Rp) configuration at one particular phosphorothioateinternucleoside linkage and the (Sp) configuration at each of theremaining phosphorothioate internucleoside linkages.

Embodiment 48. The chirally enriched population of embodiment 42 orembodiment 45 wherein the population is enriched for modifiedoligonucleotides having at least 3 contiguous phosphorothioateinternucleoside linkages in the Sp, Sp, and Rp configurations, in the 5′to 3′ direction.

Embodiment 49. The chirally enriched population of embodiment 42 orembodiment 45 wherein the population is enriched for modifiedoligonucleotides having at least 3 contiguous phosphorothioateinternucleoside linkages in the Sp, Sp, and Rp configurations, in the 5′to 3′ direction.

Embodiment 50. A chirally enriched population of oligomeric compounds ofany of embodiments 1-32, wherein all of the phosphorothioateinternucleoside linkages of the modified oligonucleotide arestereorandom.

Embodiment 51. A pharmaceutical composition comprising the modifiedoligonucleotide of any of embodiments 36-40 and a pharmaceuticallyacceptable diluent or carrier.

Embodiment 52. The pharmaceutical composition of embodiment 51, whereinthe pharmaceutically acceptable diluent is artificial cerebrospinalfluid.

Embodiment 53. The pharmaceutical composition of embodiment 50, whereinthe pharmaceutical composition consists essentially of the modifiedoligonucleotide and artificial cerebrospinal fluid.

Embodiment 54. A method comprising administering to an animal apharmaceutical composition of any of embodiments 35 or 51-53.

Embodiment 55. A method of treating a disease associated with SNCAcomprising administering to an individual having or at risk fordeveloping a disease associated with SNCA a therapeutically effectiveamount of a pharmaceutical composition according to any of embodiments35 or 51-53; and thereby treating the disease associated with SNCA.

Embodiment 56. The method of embodiment 55, wherein the diseaseassociated with SNCA is a neurodegenerative disease.

Embodiment 57. The method of embodiment 56, wherein theneurodegenerative disease is any of Parkinson's disease, dementia withLewy bodies, diffuse Lewy body disease, pure autonomic failure, multiplesystem atrophy, neuronopathic Gaucher's disease and Alzheimer's disease.

Embodiment 58. The method of embodiment 56, wherein at least one symptomor hallmark of the neurodegenerative disease is ameliorated.

Embodiment 59. The method of embodiment 58, wherein the symptom orhallmark is any of motor dysfunction, aggregation of alpha-synuclein,neurodegeneration, cognitive decline and dementia.

I. Certain Oligonucleotides

In certain embodiments, provided herein are oligomeric compoundscomprising oligonucleotides, which consist of linked nucleosides.Oligonucleotides may be unmodified oligonucleotides (RNA or DNA) or maybe modified oligonucleotides. Modified oligonucleotides comprise atleast one modification relative to unmodified RNA or DNA. That is,modified oligonucleotides comprise at least one modified nucleoside(comprising a modified sugar moiety and/or a modified nucleobase) and/orat least one modified internucleoside linkage.

A. Certain Modified Nucleosides

Modified nucleosides comprise a modified sugar moiety or a modifiednucleobase or both a modified sugar moiety and a modified nucleobase.

1. Certain Sugar Moieties

In certain embodiments, modified sugar moieties are non-bicyclicmodified sugar moieties. In certain embodiments, modified sugar moietiesare bicyclic or tricyclic sugar moieties. In certain embodiments,modified sugar moieties are sugar surrogates. Such sugar surrogates maycomprise one or more substitutions corresponding to those of other typesof modified sugar moieties.

In certain embodiments, modified sugar moieties are non-bicyclicmodified sugar moieties comprising a furanosyl ring with one or moresubstituent groups none of which bridges two atoms of the fumnosyl ringto form a bicyclic structure. Such non bridging substituents may be atany position of the furanosyl, including but not limited to substituentsat the 2′, 4′, and/or 5′ positions. In certain embodiments one or morenon-bridging substituent of non-bicyclic modified sugar moieties isbranched. Examples of 2′-substituent groups suitable for non-bicyclicmodified sugar moieties include but are not limited to: 2′-F, 2′-OCH₃(“OMe” or “O-methyl”), and 2′-O(CH₂)₂OCH₃ (“MOE”). In certainembodiments, 2′-substituent groups are selected from among: halo, allyl,amino, azido, SH, CN, OCN, CF₃, OCF₃, O—C₁-C₁₀alkoxy, O—C₁-C₁₀substituted alkoxy, O—C₁-C₁₀ alkyl, O—C₁-C₁₀ substituted alkyl, S-alkyl,N(R_(m))-alkyl, O-alkenyl, S-alkenyl, N(R_(m))-alkenyl, O-alkynyl,S-alkynyl, N(R_(m))-alkynyl, O-alkylenyl-O-alkyl, alkynyl, alkaryl,aralkyl, O-alkaryl, O-aralkyl, O(CH₂)₂SCH₃, O(CH₂)₂ON(R_(m))(R_(n)) orOCH₂C(═O)—N(R_(m))(R_(n)), where each R_(m) and R_(n) is, independently,H, an amino protecting group, or substituted or unsubstituted C₁-C₁₀alkyl, and the 2′-substituent groups described in Cook et al., U.S. Pat.No. 6,531,584; Cook et al., U.S. Pat. No. 5,859,221; and Cook et al.,U.S. Pat. No. 6,005,087. Certain embodiments of these 2′-substituentgroups can be further substituted with one or more substituent groupsindependently selected from among: hydroxyl, amino, alkoxy, carboxy,benzyl, phenyl, nitro (NO₂), thiol, thioalkoxy, thioalkyl, halogen,alkyl, aryl, alkenyl and alkynyl. Examples of 4′-substituent groupssuitable for non-bicyclic modified sugar moieties include but are notlimited to alkoxy (e.g., methoxy), alkyl, and those described inManoharan et al., WO 2015/106128. Examples of 5′-substituent groupssuitable for non-bicyclic modified sugar moieties include but are notlimited to: 5′-methyl (R or S), 5′-vinyl, and 5′-methoxy. In certainembodiments, non-bicyclic modified sugar moieties comprise more than onenon-bridging sugar substituent, for example, 2′-F-5′-methyl sugarmoieties and the modified sugar moieties and modified nucleosidesdescribed in Migawa et al., WO 2008/101157 and Rajeev et al.,US2013/0203836.).

In certain embodiments, a 2′-substituted non-bicyclic modifiednucleoside comprises a sugar moiety comprising a non-bridging2′-substituent group selected from: F, NH₂, N₃, OCF₃, OCH₃, O(CH₂)₃NH₂,CH₂CH═CH₂, OCH₂CH═CH₂, OCH₂CH₂OCH₃, O(CH₂)₂SCH₃,O(CH₂)₂ON(R_(m))(R_(n)), O(CH₂)₂O(CH₂)₂N(CH₃)₂, and N-substitutedacetamide (OCH₂C(═O)—N(R_(m))(R_(n))), where each R_(m) and R_(n) is,independently, H, an amino protecting group, or substituted orunsubstituted C₁-C₁₀ alkyl.

In certain embodiments, a 2′-substituted nucleoside non-bicyclicmodified nucleoside comprises a sugar moiety comprising a non-bridging2′-substituent group selected from: F, OCF₃, OCH₃, OCH₂CH₂OCH₃,O(CH₂)₂SCH₃, O(CH₂)₂ON(CH₃)₂, O(CH₂)₂O(CH₂)₂N(CH₃)₂, andOCH₂C(═O)—N(H)CH₃ (“NMA”).

In certain embodiments, a 2′-substituted non-bicyclic modifiednucleoside comprises a sugar moiety comprising a non-bridging2′-substituent group selected from: F, OCH₃, and OCH₂CH₂OCH₃.

Certain modified sugar moieties comprise a substituent that bridges twoatoms of the furanosyl ring to form a second ring, resulting in abicyclic sugar moiety. In certain such embodiments, the bicyclic sugarmoiety comprises a bridge between the 4′ and the 2′ furanose ring atoms.Examples of such 4′ to 2′ bridging sugar substituents include but arenot limited to: 4′-CH₂-2′, 4′-(CH₂)₂-2′, 4′-(CH₂)₃-2′, 4′-CH₂—O-2′(“LNA”), 4′-CH₂—S-2′, 4′-(CH₂)₂—O-2′ (“ENA”), 4′-CH(CH₃)—O-2′ (referredto as “constrained ethyl” or “cEt”), 4′-CH₂—N(R)-2′, 4′-CH(CH₂OCH₃)—O-2′(“constrained MOE” or “cMOE”) and analogs thereof (see, e.g., Seth etal., U.S. Pat. No. 7,399,845, Bhat et al., U.S. Pat. No. 7,569,686,Swayze et al., U.S. Pat. No. 7,741,457, and Swayze et al., U.S. Pat. No.8,022,193), 4′-C(CH₃)(CH₃)—O-2′ and analogs thereof (see, e.g., Seth etal., U.S. Pat. No. 8,278,283), 4′-CH₂—N(OCH₃)-2′ and analogs thereof(see, e.g., Prakash et al., U.S. Pat. No. 8,278,425), 4′-CH₂—O—N(CH₃)-2′(see, e.g., Allerson et al., U.S. Pat. No. 7,696,345 and Allerson etal., U.S. Pat. No. 8,124,745), 4′-CH₂—C(H)(CH₃)-2′ (see, e.g., Zhou, etal., J. Org. Chem., 2009, 74, 118-134), 4′-CH₂—C(═CH₂)-2′ and analogsthereof (see e.g., Seth et al., U.S. Pat. No. 8,278,426),4′-C(R_(a)R_(b))—N(R)—O-2′, 4′-C(R_(a)R_(b))—O—N(R)-2′,4′-CH₂—O—N(R)-2′, and 4′-CH₂—N(R)—O- 2′, wherein each R, R_(a), andR_(b) is, independently, H, a protecting group, or C₁-C₁₂ alkyl (see,e.g. Imanishi et al., U.S. Pat. No. 7,427,672).

In certain embodiments, such 4′ to 2′ bridges independently comprisefrom 1 to 4 linked groups independently selected from:—[C(R_(a))(R_(b))]_(n)—, —[C(R_(a))(R_(b))]_(n)—O—, —C(R_(a))═C(R_(b))—,—C(R_(a))═N—, —C(═NR_(a))—, —C(═O)—, —C(═S)—, —O—, —Si(R_(a))₂—,—S(═O)_(x)—, and —N(R_(a))—;

wherein:

x is 0, 1, or 2;

n is 1, 2, 3, or 4;

each R_(a) and R_(b) is, independently, H, a protecting group, hydroxyl,C₁-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, substitutedC₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C₁₂ alkynyl, C₅-C₂₀ aryl,substituted C₅-C₂₀ aryl, heterocycle radical, substituted heterocycleradical, heteroaryl, substituted heteroaryl, C₅-C₇ alicyclic radical,substituted C₅-C₇ alicyclic radical, halogen, OJ₁, NJ₁J₂, SJ₁, N₃,COOJ₁, acyl (C(═O)—H), substituted acyl, CN, sulfonyl (S(═O)₂-J₁), orsulfoxyl (S(═O)-J₁); and

each J₁ and J₂ is, independently, H, C₁-C₁₂ alkyl, substituted C₁-C₁₂alkyl, C₂-C₁₂ alkenyl, substituted C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl,substituted C₂-C₁₂ alkynyl, C₅-C₂₀ aryl, substituted C₅-C₂₀ aryl, acyl(C(═O)—H), substituted acyl, a heterocycle radical, a substitutedheterocycle radical, C₁-C₁₂ aminoalkyl, substituted C₁-C₁₂ aminoalkyl,or a protecting group.

Additional bicyclic sugar moieties are known in the art, see, forexample: Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443,Albaek et al., J. Org. Chem., 2006, 71, 7731-7740, Singh et al., Chem.Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54,3607-3630; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222;Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastava et al.,J. Am. Chem. Soc., 20017, 129, 8362-8379; Wengel et a., U.S. Pat. No.7,053,207; Imanishi et al., U.S. Pat. No. 6,268,490; Imanishi et al.U.S. Pat. No. 6,770,748; Imanishi et al., U.S. RE44,779; Wengel et al.,U.S. Pat. No. 6,794,499; Wengel et al., U.S. Pat. No. 6,670,461; Wengelet al., U.S. Pat. No. 7,034,133; Wengel et al., U.S. Pat. No. 8,080,644;Wengel et al., U.S. Pat. No. 8,034,909; Wengel et al., U.S. Pat. No.8,153,365; Wengel et al., U.S. Pat. No. 7,572,582; and Ramasamy et al.,U.S. Pat. No. 6,525,191; Torsten et al., WO 2004/106356; Wengel et al.,WO 1999/014226; Seth et al., WO 2007/134181; Seth et al., U.S. Pat. No.7,547,684; Seth et al., U.S. Pat. No. 7,666,854; Seth et al., U.S. Pat.No. 8,088,746; Seth et al., U.S. Pat. No. 7,750,131; Seth et al., U.S.Pat. No. 8,030,467; Seth et al., U.S. Pat. No. 8,268,980; Seth et al.,U.S. Pat. No. 8,546,556; Seth et al., U.S. Pat. No. 8,530,640; Migawa etal., U.S. Pat. No. 9,012,421; Seth et al., U.S. Pat. No. 8,501,805; andU.S. Patent Publication Nos. Allerson et al., US2008/0039618 and Migawaet al., US2015/0191727.

In certain embodiments, bicyclic sugar moieties and nucleosidesincorporating such bicyclic sugar moieties are further defined byisomeric configuration. For example, an LNA nucleoside (describedherein) may be in the α-L configuration or in the β-D configuration.

α-L-methyleneoxy (4′-CH₂—O-2′) or α-L-LNA bicyclic nucleosides have beenincorporated into oligonucleotides that showed antisense activity(Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372). Herein,general descriptions of bicyclic nucleosides include both isomericconfigurations. When the positions of specific bicyclic nucleosides(e.g., LNA or cEt) are identified in exemplified embodiments herein,they are in the β-D configuration, unless otherwise specified.

In certain embodiments, modified sugar moieties comprise one or morenon-bridging sugar substituent and one or more bridging sugarsubstituent (e.g., 5′-substituted and 4′-2′ bridged sugars).

In certain embodiments, modified sugar moieties are sugar surrogates. Incertain such embodiments, the oxygen atom of the sugar moiety isreplaced, e.g., with a sulfur, carbon or nitrogen atom. In certain suchembodiments, such modified sugar moieties also comprise bridging and/ornon-bridging substituents as described herein. For example, certainsugar surrogates comprise a 4′-sulfur atom and a substitution at the2′-position (see, e.g., Bhat et al., U.S. Pat. No. 7,875,733 and Bhat etal., U.S. Pat. No. 7,939,677) and/or the 5′ position.

In certain embodiments, sugar surrogates comprise rings having otherthan 5 atoms. For example, in certain embodiments, a sugar surrogatecomprises a six-membered tetrahydropyran (“THP”). Such tetrahydropyransmay be further modified or substituted. Nucleosides comprising suchmodified tetrahydropyrans include but are not limited to hexitol nucleicacid (“HNA”), anitol nucleic acid (“ANA”), manitol nucleic acid (“MNA”)(see, e.g., Leumann, C J. Bioorg. & Med. Chem. 2002, 10, 841-854),fluoro HNA:

(“F-HNA”, see e.g. Swayze et al., U.S. Pat. No. 8,088,904; Swayze etal., U.S. Pat. No. 8,440,803; Swayze et al., U.S. Pat. No. 8,796,437;and Swayze et al., U.S. Pat. No. 9,005,906; F-HNA can also be referredto as a F-THP or 3′-fluoro tetrahydropyran), and nucleosides comprisingadditional modified THP compounds having the formula:

wherein, independently, for each of said modified THP nucleoside:

Bx is a nucleobase moiety;

T₃ and T₄ are each, independently, an internucleoside linking grouplinking the modified THP nucleoside to the remainder of anoligonucleotide or one of T₃ and T₄ is an internucleoside linking grouplinking the modified THP nucleoside to the remainder of anoligonucleotide and the other of T₃ and T₄ is H, a hydroxyl protectinggroup, a linked conjugate group, or a 5′ or 3′-terminal group;

q₁, q₂, q₃, q₄, q₅, q₆ and q₇ are each, independently, H, C₁-C₆ alkyl,substituted C₁-C₆ alkyl, C₂-C₆ alkenyl, substituted C₂-C₆ alkenyl, C₂-C₆alkynyl, or substituted C₂-C₆ alkynyl; and

each of R₁ and R₂ is independently selected from among: hydrogen,halogen, substituted or unsubstituted alkoxy, NJ₁J₂, SJ₁, N₃, OC(═X)J₁,OC(═X)NJ₁J₂, NJ₃C(═X)NJ₁J₂, and CN, wherein X is O, S or NJ₁, and eachJ₁, J₂, and J₃ is, independently, H or C₁-C₆ alkyl.

In certain embodiments, modified THP nucleosides are provided whereinq₁, q₂, q₃, q₄, q₅, q₆ and q₇ are each H.

In certain embodiments, at least one of q₁, q₂, q₃, q₄, q₅, q₆ and q₇ isother than H. In certain embodiments, at least one of q₁, q₂, q₃, q₄,q₅, q₆ and q₇ is methyl. In certain embodiments, modified THPnucleosides are provided wherein one of R₁ and R₂ is F. In certainembodiments, R₁ is F and R₂ is H, in certain embodiments, R₁ is methoxyand R₂ is H, and in certain embodiments, R₁ is methoxyethoxy and R₂ isH.

In certain embodiments, sugar surrogates comprise rings having more than5 atoms and more than one heteroatom. For example, nucleosidescomprising morpholino sugar moieties and their use in oligonucleotideshave been reported (see, e.g., Braasch et al., Biochemistry, 2002, 41,4503-4510 and Summerton et al., U.S. Pat. No. 5,698,685; Summerton etal., U.S. Pat. No. 5,166,315; Summerton et al., U.S. Pat. No. 5,185,444;and Summerton et al., U.S. Pat. No. 5,034,506). As used here, the term“morpholino” means a sugar surrogate having the following structure:

In certain embodiments, morpholinos may be modified, for example byadding or altering various substituent groups from the above morpholinostructure. Such sugar surrogates are referred to herein as “modifiedmorpholinos.”

In certain embodiments, sugar surrogates comprise acyclic moieties.Examples of nucleosides and oligonucleotides comprising such acyclicsugar surrogates include but are not limited to: peptide nucleic acid(“PNA”), acyclic butyl nucleic acid (see, e.g., Kumar et al., Org.Biomol. Chem., 2013, 11, 5853-5865), and nucleosides andoligonucleotides described in Manoharan et al., WO2011/133876.

Many other bicyclic and tricyclic sugar and sugar surrogate ring systemsare known in the art that can be used in modified nucleosides).

2. Certain Modified Nucleobases

In certain embodiments, modified oligonucleotides comprise one or morenucleoside comprising an unmodified nucleobase. In certain embodiments,modified oligonucleotides comprise one or more nucleoside comprising amodified nucleobase. In certain embodiments, modified oligonucleotidescomprise one or more nucleoside that does not comprise a nucleobase,referred to as an abasic nucleoside.

In certain embodiments, modified nucleobases are selected from:5-substituted pyrimidines, 6-azapyrimidines, alkyl or alkynylsubstituted pyrimidines, alkyl substituted purines, and N-2, N-6 and 0-6substituted purines. In certain embodiments, modified nucleobases areselected from: 2-aminopropyladenine, 5-hydroxymethyl cytosine, xanthine,hypoxanthine, 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine,2-propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine,5-propynyl (—C≡C—CH₃) uracil, 5-propynylcytosine, 6-azouracil,6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil),4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-azaand other 8-substituted purines, 5-halo, particularly 5-bromo,5-trifluoromethyl, 5-halouracil, and 5-halocytosine, 7-methylguanine,7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine,7-deazaadenine, 3-deazaguanine, 3-deazaadenine, 6-N-benzoyladenine,2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl4-N-benzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases,hydrophobic bases, promiscuous bases, size-expanded bases, andfluorinated bases. Further modified nucleobases include tricyclicpyrimidines, such as 1,3-diazaphenoxazine-2-one,1,3-diazaphenothiazine-2-one and9-(2-aminoethoxy)-1,3-diazaphenoxazine-2-one (G-clamp) Modifiednucleobases may also include those in which the purine or pyrimidinebase is replaced with other heterocycles, for example 7-deaza-adenine,7-deazaguanosine, 2-aminopyridine and 2-pyridone. Further nucleobasesinclude those disclosed in Merigan et al., U.S. Pat. No. 3,687,808,those disclosed in The Concise Encyclopedia Of Polymer Science AndEngineering, Kroschwitz, J. I., Ed., John Wiley & Sons, 1990, 858-859;Englisch et al., Angewandte Chemie, International Edition, 1991, 30,613; Sanghvi, Y. S., Chapter 15, Antisense Research and Applications,Crooke, S. T. and Lebleu, B., Eds., CRC Press, 1993, 273-288; and thosedisclosed in Chapters 6 and 15, Antisense Drug Technology, Crooke S. T.,Ed., CRC Press, 2008, 163-166 and 442-443.

Publications that teach the preparation of certain of the above notedmodified nucleobases as well as other modified nucleobases includewithout limitation, Manohara et al., US2003/0158403; Manoharan et al.,US2003/0175906; Dinh et al., U.S. Pat. No. 4,845,205; Spielvogel et al.,U.S. Pat. No. 5,130,302; Rogers et al., U.S. Pat. No. 5,134,066;Bischofberger et al., U.S. Pat. No. 5,175,273; Urdea et al., U.S. Pat.No. 5,367,066; Benner et al., U.S. Pat. No. 5,432,272; Matteucci et al.,U.S. Pat. No. 5,434,257; Gmeiner et al., U.S. Pat. No. 5,457,187; Cooket al., U.S. Pat. No. 5,459,255; Froehler et al., U.S. Pat. No.5,484,908; Matteucci et al., U.S. Pat. No. 5,502,177; Hawkins et al.,U.S. Pat. No. 5,525,711; Haralambidis et al., U.S. Pat. No. 5,552,540;Cook et al., U.S. Pat. No. 5,587,469; Froehler et al., U.S. Pat. No.5,594,121; Switzer et al., U.S. Pat. No. 5,596,091; Cook et al., U.S.Pat. No. 5,614,617; Froehler et al., U.S. Pat. No. 5,645,985; Cook etal., U.S. Pat. No. 5,681,941; Cook et al., U.S. Pat. No. 5,811,534; Cooket al., U.S. Pat. No. 5,750,692; Cook et al., U.S. Pat. No. 5,948,903;Cook et al., U.S. Pat. No. 5,587,470; Cook et al., U.S. Pat. No.5,457,191; Matteucci et al., U.S. Pat. No. 5,763,588; Froehler et al.,U.S. Pat. No. 5,830,653; Cook et al., U.S. Pat. No. 5,808,027; Cook etal., 6,166,199; and Matteucci et al., U.S. Pat. No. 6,005,096.

3. Certain Modified Internucleoside Linkages

In certain embodiments, nucleosides of modified oligonucleotides may belinked together using any internucleoside linkage. The two main classesof internucleoside linking groups are defined by the presence or absenceof a phosphorus atom. Representative phosphorus-containinginternucleoside linkages include but are not limited to phosphates,which contain a phosphodiester bond (“P═O”) (also referred to asunmodified or naturally occurring linkages), phosphotriesters,methylphosphonates, phosphoramidates, and phosphorothioates (“P═S”), andphosphorodithioates (“HS-P═S”). Representative non-phosphorus containinginternucleoside linking groups include but are not limited tomethylenemethylimino (—CH₂—N(CH₃)—O—CH₂—), thiodiester, thionocarbamate(—O—C(═O)(NH)—S—); siloxane (—O—SiH₂—O—); and N,N′-dimethylhydrazine(—CH₂—N(CH₃)—N(CH₃)—). Modified internucleoside linkages, compared tonaturally occurring phosphate linkages, can be used to alter, typicallyincrease, nuclease resistance of the oligonucleotide. In certainembodiments, internucleoside linkages having a chiral atom can beprepared as a racemic mixture, or as separate enantiomers. Methods ofpreparation of phosphorous-containing and non-phosphorous-containinginternucleoside linkages are well known to those skilled in the art.

Representative internucleoside linkages having a chiral center includebut are not limited to alkylphosphonates and phosphorothioates. Modifiedoligonucleotides comprising internucleoside linkages having a chiralcenter can be prepared as populations of modified oligonucleotidescomprising stereorandom internucleoside linkages, or as populations ofmodified oligonucleotides comprising phosphorothioate linkages inparticular stereochemical configurations. In certain embodiments,populations of modified oligonucleotides comprise phosphorothioateinternucleoside linkages wherein all of the phosphorothioateinternucleoside linkages are stereorandom. Such modifiedoligonucleotides can be generated using synthetic methods that result inrandom selection of the stereochemical configuration of eachphosphorothioate linkage. Nonetheless, as is well understood by those ofskill in the art, each individual phosphorothioate of each individualoligonucleotide molecule has a defined stereoconfiguration. In certainembodiments, populations of modified oligonucleotides are enriched formodified oligonucleotides comprising one or more particularphosphorothioate internucleoside linkages in a particular, independentlyselected stereochemical configuration. In certain embodiments, theparticular configuration of the particular phosphorothioate linkage ispresent in at least 65% of the molecules in the population. In certainembodiments, the particular configuration of the particularphosphorothioate linkage is present in at least 70% of the molecules inthe population. In certain embodiments, the particular configuration ofthe particular phosphorothioate linkage is present in at least 80% ofthe molecules in the population. In certain embodiments, the particularconfiguration of the particular phosphorothioate linkage is present inat least 90% of the molecules in the population. In certain embodiments,the particular configuration of the particular phosphorothioate linkageis present in at least 99% of the molecules in the population. Suchchirally enriched populations of modified oligonucleotides can begenerated using synthetic methods known in the art, e.g., methodsdescribed in Oka et al., JACS 125, 8307 (2003), Wan et al. Nuc. Acid.Res. 42, 13456 (2014), and WO 2017/015555. In certain embodiments, apopulation of modified oligonucleotides is enriched for modifiedoligonucleotides having at least one indicated phosphorothioate in the(Sp) configuration. In certain embodiments, a population of modifiedoligonucleotides is enriched for modified oligonucleotides having atleast one phosphorothioate in the (Rp) configuration. In certainembodiments, modified oligonucleotides comprising (Rp) and/or (Sp)phosphorothioates comprise one or more of the following formulas,respectively, wherein “B” indicates a nucleobase:

Unless otherwise indicated, chiral internucleoside linkages of modifiedoligonucleotides described herein can be stereorandom or in a particularstereochemical configuration.

Neutral internucleoside linkages include, without limitation,phosphotriesters, methylphosphonates, MMI (3′-CH₂—N(CH₃)—O-5′), amide-3(3′-CH₂—C(═O)—N(H)-5′), amide-4 (3′-CH₂—N(H)-C(═O)-5′), formacetal(3′-O—CH₂—O-5′), methoxypropyl, and thioformacetal (3′-S—CH₂—O-5′).Further neutral internucleoside linkages include nonionic linkagescomprising siloxane (dialkylsiloxane), carboxylate ester, carboxamide,sulfide, sulfonate ester and amides (See for example: CarbohydrateModifications in Antisense Research; Y. S. Sanghvi and P. D. Cook, Eds.,ACS Symposium Series 580; Chapters 3 and 4, 40-65). Further neutralinternucleoside linkages include nonionic linkages comprising mixed N,O, S and CH₂ component parts.

B. Certain Motifs

In certain embodiments, modified oligonucleotides comprise one or moremodified nucleosides comprising a modified sugar moiety. In certainembodiments, modified oligonucleotides comprise one or more modifiednucleosides comprising a modified nucleobase. In certain embodiments,modified oligonucleotides comprise one or more modified internucleosidelinkage. In such embodiments, the modified, unmodified, and differentlymodified sugar moieties, nucleobases, and/or internucleoside linkages ofa modified oligonucleotide define a pattern or motif. In certainembodiments, the patterns of sugar moieties, nucleobases, andinternucleoside linkages are each independent of one another. Thus, amodified oligonucleotide may be described by its sugar motif, nucleobasemotif and/or internucleoside linkage motif (as used herein, nucleobasemotif describes the modifications to the nucleobases independent of thesequence of nucleobases).

1. Certain Sugar Motifs

In certain embodiments, oligonucleotides comprise one or more type ofmodified sugar and/or unmodified sugar moiety arranged along theoligonucleotide or region thereof in a defined pattern or sugar motif.In certain instances, such sugar motifs include but are not limited toany of the sugar modifications discussed herein.

In certain embodiments, modified oligonucleotides comprise or consist ofa region having a gapmer motif, which is defined by two external regionsor “wings” and a central or internal region or “gap.” The three regionsof a gapmer motif (the 5′-wing, the gap, and the 3′-wing) form acontiguous sequence of nucleosides wherein at least some of the sugarmoieties of the nucleosides of each of the wings differ from at leastsome of the sugar moieties of the nucleosides of the gap. Specifically,at least the sugar moieties of the nucleosides of each wing that areclosest to the gap (the 3′-most nucleoside of the 5′-wing and the5′-most nucleoside of the 3′-wing) differ from the sugar moiety of theneighboring gap nucleosides, thus defining the boundary between thewings and the gap (i.e., the wing/gap junction). In certain embodiments,the sugar moieties within the gap are the same as one another. Incertain embodiments, the gap includes one or more nucleoside having asugar moiety that differs from the sugar moiety of one or more othernucleosides of the gap. In certain embodiments, the sugar motifs of thetwo wings are the same as one another (symmetric gapmer). In certainembodiments, the sugar motif of the 5′-wing differs from the sugar motifof the 3′-wing (asymmetric gapmer).

In certain embodiments, the wings of a gapmer comprise 1-5 nucleosides.In certain embodiments, each nucleoside of each wing of a gapmer is amodified nucleoside. In certain embodiments, at least one nucleoside ofeach wing of a gapmer is a modified nucleoside. In certain embodiments,at least two nucleosides of each wing of a gapmer are modifiednucleosides. In certain embodiments, at least three nucleosides of eachwing of a gapmer are modified nucleosides. In certain embodiments, atleast four nucleosides of each wing of a gapmer are modifiednucleosides.

In certain embodiments, the gap of a gapmer comprises 7-12 nucleosides.In certain embodiments, each nucleoside of the gap of a gapmer is anunmodified 2′-deoxy nucleoside.

In certain embodiments, the gapmer is a deoxy gapmer. In embodiments,the nucleosides on the gap side of each wing/gap junction are unmodified2′-deoxy nucleosides and the nucleosides on the wing sides of eachwing/gap junction are modified nucleosides. In certain embodiments, eachnucleoside of the gap is an unmodified 2′-deoxy nucleoside. In certainembodiments, each nucleoside of each wing of a gapmer is a modifiednucleoside.

In certain embodiments, modified oligonucleotides comprise or consist ofa region having a fully modified sugar motif. In such embodiments, eachnucleoside of the fully modified region of the modified oligonucleotidecomprises a modified sugar moiety. In certain embodiments, eachnucleoside of the entire modified oligonucleotide comprises a modifiedsugar moiety. In certain embodiments, modified oligonucleotides compriseor consist of a region having a fully modified sugar motif, wherein eachnucleoside within the fully modified region comprises the same modifiedsugar moiety, referred to herein as a uniformly modified sugar motif. Incertain embodiments, a fully modified oligonucleotide is a uniformlymodified oligonucleotide. In certain embodiments, each nucleoside of auniformly modified comprises the same 2′-modification.

Herein, the lengths (number of nucleosides) of the three regions of agapmer may be provided using the notation [# of nucleosides in the5′-wing]-[# of nucleosides in the gap]-[# of nucleosides in the3′-wing]. Thus, a 5-10-5 gapmer consists of 5 linked nucleosides in eachwing and 10 linked nucleosides in the gap. Where such nomenclature isfollowed by a specific modification, that modification is themodification in each sugar moiety of each wing and the gap nucleosidescomprise unmodified deoxynucleosides sugars. Thus, a 5-10-5 MOE gapmerconsists of 5 linked MOE modified nucleosides in the 5′-wing, 10 linkeddeoxynucleosides in the gap, and 5 linked MOE nucleosides in the3′-wing.

In certain embodiments, modified oligonucleotides are 5-10-5 MOEgapmers. In certain embodiments, modified oligonucleotides are 3-10-3BNA gapmers. In certain embodiments, modified oligonucleotides are3-10-3 cEt gapmers. In certain embodiments, modified oligonucleotidesare 3-10-3 LNA gapmers.

2. Certain Nucleobase Motifs

In certain embodiments, oligonucleotides comprise modified and/orunmodified nucleobases arranged along the oligonucleotide or regionthereof in a defined pattern or motif. In certain embodiments, eachnucleobase is modified. In certain embodiments, none of the nucleobasesare modified. In certain embodiments, each purine or each pyrimidine ismodified. In certain embodiments, each adenine is modified. In certainembodiments, each guanine is modified. In certain embodiments, eachthymine is modified. In certain embodiments, each uracil is modified. Incertain embodiments, each cytosine is modified. In certain embodiments,some or all of the cytosine nucleobases in a modified oligonucleotideare 5-methyl cytosines. In certain embodiments, all of the cytosinenucleobases are 5-methyl cytosines and all of the other nucleobases ofthe modified oligonucleotide are unmodified nucleobases.

In certain embodiments, modified oligonucleotides comprise a block ofmodified nucleobases. In certain such embodiments, the block is at the3′-end of the oligonucleotide. In certain embodiments the block iswithin 3 nucleosides of the 3′-end of the oligonucleotide. In certainembodiments, the block is at the 5′-end of the oligonucleotide. Incertain embodiments the block is within 3 nucleosides of the 5′-end ofthe oligonucleotide.

In certain embodiments, oligonucleotides having a gapmer motif comprisea nucleoside comprising a modified nucleobase. In certain suchembodiments, one nucleoside comprising a modified nucleobase is in thecentral gap of an oligonucleotide having a gapmer motif. In certain suchembodiments, the sugar moiety of said nucleoside is a 2′-deoxyribosylmoiety. In certain embodiments, the modified nucleobase is selectedfrom: a 2-thiopyrimidine and a 5-propynepyrimidine.

3. Certain Internucleoside Linkage Motifs

In certain embodiments, oligonucleotides comprise modified and/orunmodified internucleoside linkages arranged along the oligonucleotideor region thereof in a defined pattern or motif. In certain embodiments,each internucleoside linking group is a phosphodiester internucleosidelinkage (P═O). In certain embodiments, each internucleoside linkinggroup of a modified oligonucleotide is a phosphorothioateinternucleoside linkage (P═S). In certain embodiments, eachinternucleoside linkage of a modified oligonucleotide is independentlyselected from a phosphorothioate internucleoside linkage andphosphodiester internucleoside linkage. In certain embodiments, eachphosphorothioate internucleoside linkage is independently selected froma stereorandom phosphorothioate a (Sp) phosphorothioate, and a (Rp)phosphorothioate. In certain embodiments, the sugar motif of a modifiedoligonucleotide is a gapmer and the internucleoside linkages within thegap are all modified. In certain such embodiments, some or all of theinternucleoside linkages in the wings are unmodified phosphodiesterinternucleoside linkages. In certain embodiments, the terminalinternucleoside linkages are modified. In certain embodiments, the sugarmotif of a modified oligonucleotide is a gapmer, and the internucleosidelinkage motif comprises at least one phosphodiester internucleosidelinkage in at least one wing, wherein the at least one phosphodiesterlinkage is not a terminal internucleoside linkage, and the remaininginternucleoside linkages are phosphorothioate internucleoside linkages.In certain such embodiments, all of the phosphorothioate linkages arestereorandom. In certain embodiments, all of the phosphorothioatelinkages in the wings are (Sp) phosphorothioates, and the gap comprisesat least one Sp, Sp, Rp motif. In certain embodiments, populations ofmodified oligonucleotides are enriched for modified oligonucleotidescomprising such internucleoside linkage motifs.

C. Certain Lengths

It is possible to increase or decrease the length of an oligonucleotidewithout eliminating activity. For example, in Woolf et al. (Proc. Natl.Acad. Sci. USA 89:7305-7309, 1992), a series of oligonucleotides 13-25nucleobases in length were tested for their ability to induce cleavageof a target RNA in an oocyte injection model. Oligonucleotides 25nucleobases in length with 8 or 11 mismatch bases near the ends of theoligonucleotides were able to direct specific cleavage of the targetmRNA, albeit to a lesser extent than the oligonucleotides that containedno mismatches. Similarly, target specific cleavage was achieved using 13nucleobase oligonucleotides, including those with 1 or 3 mismatches.

In certain embodiments, oligonucleotides (including modifiedoligonucleotides) can have any of a variety of ranges of lengths. Incertain embodiments, oligonucleotides consist of X to Y linkednucleosides, where X represents the fewest number of nucleosides in therange and Y represents the largest number nucleosides in the range. Incertain such embodiments, X and Y are each independently selected from8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, and 50; provided that X≤Y. For example, incertain embodiments, oligonucleotides consist of 12 to 13, 12 to 14, 12to 15, 12 to 16, 12 to 17, 12 to 18, 12 to 19, 12 to 20, 12 to 21, 12 to22, 12 to 23, 12 to 24, 12 to 25, 12 to 26, 12 to 27, 12 to 28, 12 to29, 12 to 30, 13 to 14, 13 to 15, 13 to 16, 13 to 17, 13 to 18, 13 to19, 13 to 20, 13 to 21, 13 to 22, 13 to 23, 13 to 24, 13 to 25, 13 to26, 13 to 27, 13 to 28, 13 to 29, 13 to 30, 14 to 15, 14 to 16, 14 to17, 14 to 18, 14 to 19, 14 to 20, 14 to 21, 14 to 22, 14 to 23, 14 to24, 14 to 25, 14 to 26, 14 to 27, 14 to 28, 14 to 29, 14 to 30, 15 to16, 15 to 17, 15 to 18, 15 to 19, 15 to 20, 15 to 21, 15 to 22, 15 to23, 15 to 24, 15 to 25, 15 to 26, 15 to 27, 15 to 28, 15 to 29, 15 to30, 16 to 17, 16 to 18, 16 to 19, 16 to 20, 16 to 21, 16 to 22, 16 to23, 16 to 24, 16 to 25, 16 to 26, 16 to 27, 16 to 28, 16 to 29, 16 to30, 17 to 18, 17 to 19, 17 to 20, 17 to 21, 17 to 22, 17 to 23, 17 to24, 17 to 25, 17 to 26, 17 to 27, 17 to 28, 17 to 29, 17 to 30, 18 to19, 18 to 20, 18 to 21, 18 to 22, 18 to 23, 18 to 24, 18 to 25, 18 to26, 18 to 27, 18 to 28, 18 to 29, 18 to 30, 19 to 20, 19 to 21, 19 to22, 19 to 23, 19 to 24, 19 to 25, 19 to 26, 19 to 29, 19 to 28, 19 to29, 19 to 30, 20 to 21, 20 to 22, 20 to 23, 20 to 24, 20 to 25, 20 to26, 20 to 27, 20 to 28, 20 to 29, 20 to 30, 21 to 22, 21 to 23, 21 to24, 21 to 25, 21 to 26, 21 to 27, 21 to 28, 21 to 29, 21 to 30, 22 to23, 22 to 24, 22 to 25, 22 to 26, 22 to 27, 22 to 28, 22 to 29, 22 to30, 23 to 24, 23 to 25, 23 to 26, 23 to 27, 23 to 28, 23 to 29, 23 to30, 24 to 25, 24 to 26, 24 to 27, 24 to 28, 24 to 29, 24 to 30, 25 to26, 25 to 27, 25 to 28, 25 to 29, 25 to 30, 26 to 27, 26 to 28, 26 to29, 26 to 30, 27 to 28, 27 to 29, 27 to 30, 28 to 29, 28 to 30, or 29 to30 linked nucleosides

D. Certain Modified Oligonucleotides

In certain embodiments, the above modifications (sugar, nucleobase,internucleoside linkage) are incorporated into a modifiedoligonucleotide. In certain embodiments, modified oligonucleotides arecharacterized by their modification motifs and overall lengths. Incertain embodiments, such parameters are each independent of oneanother.

Thus, unless otherwise indicated, each internucleoside linkage of anoligonucleotide having a gapmer sugar motif may be modified orunmodified and may or may not follow the gapmer modification pattern ofthe sugar modifications. For example, the internucleoside linkageswithin the wing regions of a sugar gapmer may be the same or differentfrom one another and may be the same or different from theinternucleoside linkages of the gap region of the sugar motif. Likewise,such sugar gapmer oligonucleotides may comprise one or more modifiednucleobase independent of the gapmer pattern of the sugar modifications.Unless otherwise indicated, all modifications are independent ofnucleobase sequence.

E. Certain Populations of Modified Oligonucleotides

Populations of modified oligonucleotides in which all of the modifiedoligonucleotides of the population have the same molecular formula canbe stereorandom populations or chirally enriched populations. All of thechiral centers of all of the modified oligonucleotides are stereorandomin a stereorandom population. In a chirally enriched population, atleast one particular chiral center is not stereorandom in the modifiedoligonucleotides of the population. In certain embodiments, the modifiedoligonucleotides of a chirally enriched population are enriched for β-Dribosyl sugar moieties, and all of the phosphorothioate internucleosidelinkages are stereorandom. In certain embodiments, the modifiedoligonucleotides of a chirally enriched population are enriched for bothβ-D ribosyl sugar moieties and at least one, particular phosphorothioateinternucleoside linkage in a particular stereochemical configuration.

F. Nucleobase Sequence

In certain embodiments, oligonucleotides (unmodified or modifiedoligonucleotides) are further described by their nucleobase sequence. Incertain embodiments oligonucleotides have a nucleobase sequence that iscomplementary to a second oligonucleotide or an identified referencenucleic acid, such as a target nucleic acid. In certain suchembodiments, a region of an oligonucleotide has a nucleobase sequencethat is complementary to a second oligonucleotide or an identifiedreference nucleic acid, such as a target nucleic acid. In certainembodiments, the nucleobase sequence of a region or entire length of anoligonucleotide is at least 50%, at least 60%, at least 70%, at least80%, at least 85%, at least 90%, at least 95%, or 100% complementary tothe second oligonucleotide or nucleic acid, such as a target nucleicacid.

II. Certain Oligomeric Compounds

In certain embodiments, provided herein are oligomeric compounds, whichconsist of an oligonucleotide (modified or unmodified) and optionallyone or more conjugate groups and/or terminal groups. Conjugate groupsconsist of one or more conjugate moiety and a conjugate linker whichlinks the conjugate moiety to the oligonucleotide. Conjugate groups maybe attached to either or both ends of an oligonucleotide and/or at anyinternal position. In certain embodiments, conjugate groups are attachedto the 2′-position of a nucleoside of a modified oligonucleotide. Incertain embodiments, conjugate groups that are attached to either orboth ends of an oligonucleotide are terminal groups. In certain suchembodiments, conjugate groups or terminal groups are attached at the 3′and/or 5′-end of oligonucleotides. In certain such embodiments,conjugate groups (or terminal groups) are attached at the 3′-end ofoligonucleotides. In certain embodiments, conjugate groups are attachednear the 3′-end of oligonucleotides. In certain embodiments, conjugategroups (or terminal groups) are attached at the 5′-end ofoligonucleotides. In certain embodiments, conjugate groups are attachednear the 5′-end of oligonucleotides.

Examples of terminal groups include but are not limited to conjugategroups, capping groups, phosphate moieties, protecting groups, modifiedor unmodified nucleosides, and two or more nucleosides that areindependently modified or unmodified.

A. Certain Conjugate Groups

In certain embodiments, oligonucleotides are covalently attached to oneor more conjugate groups. In certain embodiments, conjugate groupsmodify one or more properties of the attached oligonucleotide, includingbut not limited to pharmacodynamics, pharmacokinetics, stability,binding, absorption, tissue distribution, cellular distribution,cellular uptake, charge and clearance. In certain embodiments, conjugategroups impart a new property on the attached oligonucleotide, e.g.,fluorophores or reporter groups that enable detection of theoligonucleotide. Certain conjugate groups and conjugate moieties havebeen described previously, for example: cholesterol moiety (Letsinger etal., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid(Manoharan et al., Bioorg. Med. Chem. Lett., 1994, 4, 1053-1060), athioether, e.g., hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad.Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Lett.,1993, 3, 2765-2770), a thiocholesterol (Oberhauser et al., Nucl. AcidsRes., 1992, 20, 533-538), an aliphatic chain, e g., do-decan-diol orundecyl residues (Saison-Behmoaras et al., EMBO J., 1991, 10, 1111-1118;Kabanov et al., FEBS Lett., 1990, 259, 327-330; Svinarchuk et al.,Biochimie, 1993, 75, 49-54), a phospholipid, e.g.,di-hexadecyl-rac-glycerol or triethyl-ammonium1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al.,Tetrahedron Lett., 1995, 36, 3651-3654; Shea et al., Nucl. Acids Res.,1990, 18, 3777-3783), a polyamine or a polyethylene glycol chain(Manoharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), oradamantane acetic acid a palmityl moiety (Mishra et al., Biochim.Biophys. Acta, 1995, 1264, 229-237), an octadecylamine orhexylamino-cathonyl-oxycholesterol moiety (Crooke et al., J. Pharmacol.Exp. Ther., 1996, 277, 923-937), a tocopherol group (Nishina et al.,Molecular Therapy Nucleic Acids, 2015, 4, e220; and Nishina et al.,Molecular Therapy, 2008, 16, 734-740), or a GalNAc cluster (e.g.,WO2014/179620).

1. Conjugate Moieties

Conjugate moieties include, without limitation, intercalators, reportermolecules, polyamines, polyamides, peptides, carbohydrates, vitaminmoieties, polyethylene glycols, thioethers, polyethers, cholesterols,thiocholesterols, cholic acid moieties, folate, lipids, phospholipids,biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine,fluoresceins, rhodamines, coumarins, fluorophores, and dyes.

In certain embodiments, a conjugate moiety comprises an active drugsubstance, for example, aspirin, warfarin, phenylbutazone, ibuprofen,suprofen, fen-bufen, ketoprofen, (S)-(+)-pranoprofen, carprofen,dansylsarcosine, 2,3,5-triiodobenzoic acid, fingolimod, flufenamic acid,folinic acid, a benzothiadiazide, chlorothiazide, a diazepine,indo-methicin, a barbiturate, a cephalosporin, a sulfa drug, anantidiabetic, an antibacterial or an antibiotic.

2. Conjugate Linkers

Conjugate moieties are attached to oligonucleotides through conjugatelinkers. In certain oligomeric compounds, the conjugate linker is asingle chemical bond (i.e., the conjugate moiety is attached directly toan oligonucleotide through a single bond). In certain embodiments, theconjugate linker comprises a chain structure, such as a hydrocarbylchain, or an oligomer of repeating units such as ethylene glycol,nucleosides, or amino acid units.

In certain embodiments, a conjugate linker comprises one or more groupsselected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol,ether, thioether, and hydroxylamino. In certain such embodiments, theconjugate linker comprises groups selected from alkyl, amino, oxo, amideand ether groups. In certain embodiments, the conjugate linker comprisesgroups selected from alkyl and amide groups. In certain embodiments, theconjugate linker comprises groups selected from alkyl and ether groups.In certain embodiments, the conjugate linker comprises at least onephosphorus moiety. In certain embodiments, the conjugate linkercomprises at least one phosphate group. In certain embodiments, theconjugate linker includes at least one neutral linking group.

In certain embodiments, conjugate linkers, including the conjugatelinkers described above, are bifunctional linking moieties, e.g., thoseknown in the art to be useful for attaching conjugate groups to parentcompounds, such as the oligonucleotides provided herein. In general, abifunctional linking moiety comprises at least two functional groups.One of the functional groups is selected to bind to a particular site ona parent compound and the other is selected to bind to a conjugategroup. Examples of functional groups used in a bifunctional linkingmoiety include but are not limited to electrophiles for reacting withnucleophilic groups and nucleophiles for reacting with electrophilicgroups. In certain embodiments, bifunctional linking moieties compriseone or more groups selected from amino, hydroxyl, carboxylic acid,thiol, alkyl, alkenyl, and alkynyl.

Examples of conjugate linkers include but are not limited topyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) and6-aminohexanoic acid (AHEX or AHA). Other conjugate linkers include butare not limited to substituted or unsubstituted C₁-C₁₀ alkyl,substituted or unsubstituted C₂-C₁₀ alkenyl or substituted orunsubstituted C₂-C₁₀ alkynyl, wherein a nonlimiting list of preferredsubstituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl,phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl andalkynyl.

In certain embodiments, conjugate linkers comprise 1-10linker-nucleosides. In certain embodiments, conjugate linkers comprise2-5 linker-nucleosides. In certain embodiments, conjugate linkerscomprise exactly 3 linker-nucleosides. In certain embodiments, conjugatelinkers comprise the TCA motif. In certain embodiments, suchlinker-nucleosides are modified nucleosides. In certain embodiments suchlinker-nucleosides comprise a modified sugar moiety. In certainembodiments, linker-nucleosides are unmodified. In certain embodiments,linker-nucleosides comprise an optionally protected heterocyclic baseselected from a purine, substituted purine, pyrimidine or substitutedpyrimidine. In certain embodiments, a cleavable moiety is a nucleosideselected from uracil, thymine, cytosine, 4-N-benzoylcytosine, 5-methylcytosine, 4-N-benzoyl-5-methyl cytosine, adenine, 6-N-benzoyladenine,guanine and 2-N-isobutyrylguanine. It is typically desirable forlinker-nucleosides to be cleaved from the oligomeric compound after itreaches a target tissue. Accordingly, linker-nucleosides are typicallylinked to one another and to the remainder of the oligomeric compoundthrough cleavable bonds. In certain embodiments, such cleavable bondsare phosphodiester bonds.

Herein, linker-nucleosides are not considered to be part of theoligonucleotide. Accordingly, in embodiments in which an oligomericcompound comprises an oligonucleotide consisting of a specified numberor range of linked nucleosides and/or a specified percentcomplementarity to a reference nucleic acid and the oligomeric compoundalso comprises a conjugate group comprising a conjugate linkercomprising linker-nucleosides, those linker-nucleosides are not countedtoward the length of the oligonucleotide and are not used in determiningthe percent complementarity of the oligonucleotide for the referencenucleic acid. For example, an oligomeric compound may comprise (1) amodified oligonucleotide consisting of 8-30 nucleosides and (2) aconjugate group comprising 1-10 linker-nucleosides that are contiguouswith the nucleosides of the modified oligonucleotide. The total numberof contiguous linked nucleosides in such an oligomeric compound is morethan 30. Alternatively, an oligomeric compound may comprise a modifiedoligonucleotide consisting of 8-30 nucleosides and no conjugate group.The total number of contiguous linked nucleosides in such an oligomericcompound is no more than 30. Unless otherwise indicated conjugatelinkers comprise no more than 10 linker-nucleosides. In certainembodiments, conjugate linkers comprise no more than 5linker-nucleosides. In certain embodiments, conjugate linkers compriseno more than 3 linker-nucleosides. In certain embodiments, conjugatelinkers comprise no more than 2 linker-nucleosides. In certainembodiments, conjugate linkers comprise no more than 1linker-nucleoside.

In certain embodiments, it is desirable for a conjugate group to becleaved from the oligonucleotide. For example, in certain circumstancesoligomeric compounds comprising a particular conjugate moiety are bettertaken up by a particular cell type, but once the oligomeric compound hasbeen taken up, it is desirable that the conjugate group be cleaved torelease the unconjugated or parent oligonucleotide. Thus, certainconjugate linkers may comprise one or more cleavable moieties. Incertain embodiments, a cleavable moiety is a cleavable bond. In certainembodiments, a cleavable moiety is a group of atoms comprising at leastone cleavable bond. In certain embodiments, a cleavable moiety comprisesa group of atoms having one, two, three, four, or more than fourcleavable bonds. In certain embodiments, a cleavable moiety isselectively cleaved inside a cell or subcellular compartment, such as alysosome. In certain embodiments, a cleavable moiety is selectivelycleaved by endogenous enzymes, such as nucleases.

In certain embodiments, a cleavable bond is selected from among: anamide, an ester, an ether, one or both esters of a phosphodiester, aphosphate ester, a carbamate, or a disulfide. In certain embodiments, acleavable bond is one or both of the esters of a phosphodiester. Incertain embodiments, a cleavable moiety comprises a phosphate orphosphodiester. In certain embodiments, the cleavable moiety is aphosphate linkage between an oligonucleotide and a conjugate moiety orconjugate group.

In certain embodiments, a cleavable moiety comprises or consists of oneor more linker-nucleosides. In certain such embodiments, the one or morelinker-nucleosides are linked to one another and/or to the remainder ofthe oligomeric compound through cleavable bonds. In certain embodiments,such cleavable bonds are unmodified phosphodiester bonds. In certainembodiments, a cleavable moiety is 2′-deoxy nucleoside that is attachedto either the 3′ or 5′-terminal nucleoside of an oligonucleotide by aphosphate internucleoside linkage and covalently attached to theremainder of the conjugate linker or conjugate moiety by a phosphate orphosphorothioate linkage. In certain such embodiments, the cleavablemoiety is 2′-deoxyadenosine.

B. Certain Terminal Groups

In certain embodiments, oligomeric compounds comprise one or moreterminal groups. In certain such embodiments, oligomeric compoundscomprise a stabilized 5′-phophate. Stabilized 5′-phosphates include, butare not limited to 5′-phosphanates, including, but not limited to5′-vinylphosphonates. In certain embodiments, terminal groups compriseone or more abasic nucleosides and/or inverted nucleosides. In certainembodiments, terminal groups comprise one or more 2′-linked nucleosides.In certain such embodiments, the 2′-linked nucleoside is an abasicnucleoside.

III. Oligomeric Duplexes

In certain embodiments, oligomeric compounds described herein comprisean oligonucleotide, having a nucleobase sequence complementary to thatof a target nucleic acid. In certain embodiments, an oligomeric compoundis paired with a second oligomeric compound to form an oligomericduplex. Such oligomeric duplexes comprise a first oligomeric compoundhaving a region complementary to a target nucleic acid and a secondoligomeric compound having a region complementary to the firstoligomeric compound. In certain embodiments, the first oligomericcompound of an oligomeric duplex comprises or consists of (1) a modifiedor unmodified oligonucleotide and optionally a conjugate group and (2) asecond modified or unmodified oligonucleotide and optionally a conjugategroup. Either or both oligomeric compounds of an oligomeric duplex maycomprise a conjugate group. The oligonucleotides of each oligomericcompound of an oligomeric duplex may include non-complementaryoverhanging nucleosides.

IV. Antisense Activity

In certain embodiments, oligomeric compounds and oligomeric duplexes arecapable of hybridizing to a target nucleic acid, resulting in at leastone antisense activity; such oligomeric compounds and oligomericduplexes are antisense compounds. In certain embodiments, antisensecompounds have antisense activity when they reduce or inhibit the amountor activity of a target nucleic acid by 25% or more in the standard cellassay. In certain embodiments, antisense compounds selectively affectone or more target nucleic acid. Such antisense compounds comprise anucleobase sequence that hybridizes to one or more target nucleic acid,resulting in one or more desired antisense activity and does nothybridize to one or more non-target nucleic acid or does not hybridizeto one or more non-target nucleic acid in such a way that results insignificant undesired antisense activity.

In certain antisense activities, hybridization of an antisense compoundto a target nucleic acid results in recruitment of a protein thatcleaves the target nucleic acid. For example, certain antisensecompounds result in RNase H mediated cleavage of the target nucleicacid. RNase H is a cellular endonuclease that cleaves the RNA strand ofan RNA:DNA duplex. The DNA in such an RNA:DNA duplex need not beunmodified DNA. In certain embodiments, described herein are antisensecompounds that are sufficiently “DNA-like” to elicit RNase H activity.In certain embodiments, one or more non-DNA-like nucleoside in the gapof a gapmer is tolerated.

In certain antisense activities, an antisense compound or a portion ofan antisense compound is loaded into an RNA-induced silencing complex(RISC), ultimately resulting in cleavage of the target nucleic acid. Forexample, certain antisense compounds result in cleavage of the targetnucleic acid by Argonaute Antisense compounds that are loaded into RISCare RNAi compounds. RNAi compounds may be double-stranded (siRNA) orsingle-stranded (ssRNA).

In certain embodiments, hybridization of an antisense compound to atarget nucleic acid does not result in recruitment of a protein thatcleaves that target nucleic acid. In certain embodiments, hybridizationof the antisense compound to the target nucleic acid results inalteration of splicing of the target nucleic acid. In certainembodiments, hybridization of an antisense compound to a target nucleicacid results in inhibition of a binding interaction between the targetnucleic acid and a protein or other nucleic acid. In certainembodiments, hybridization of an antisense compound to a target nucleicacid results in alteration of translation of the target nucleic acid.

Antisense activities may be observed directly or indirectly. In certainembodiments, observation or detection of an antisense activity involvesobservation or detection of a change in an amount of a target nucleicacid or protein encoded by such target nucleic acid, a change in theratio of splice variants of a nucleic acid or protein and/or aphenotypic change in a cell or animal

V. Certain Target Nucleic Acids

In certain embodiments, oligomeric compounds comprise or consist of anoligonucleotide comprising a region that is complementary to a targetnucleic acid. In certain embodiments, the target nucleic acid is anendogenous RNA molecule. In certain embodiments, the target nucleic acidencodes a protein. In certain such embodiments, the target nucleic acidis selected from: a mature mRNA and a pre-mRNA, including intronic,exonic and untranslated regions. In certain embodiments, the target RNAis a mature mRNA. In certain embodiments, the target nucleic acid is apre-mRNA. In certain such embodiments, the target region is entirelywithin an intron. In certain embodiments, the target region spans anintron/exon junction. In certain embodiments, the target region is atleast 50% within an intron. In certain embodiments, the target nucleicacid is the RNA transcriptional product of a retrogene. In certainembodiments, the target nucleic acid is a non-coding RNA. In certainsuch embodiments, the target non-coding RNA is selected from: a longnon-coding RNA, a short non-coding RNA, an intronic RNA molecule.

A. Complementarity/Mismatches to the Target Nucleic Acid

It is possible to introduce mismatch bases without eliminating activity.For example, Gautschi et al (J. Natl. Cancer Inst. 93:463-471, March2001) demonstrated the ability of an oligonucleotide having 100%complementarity to the bcl-2 mRNA and having 3 mismatches to the bcl-xLmRNA to reduce the expression of both bcl-2 and bc1-xL in vitro and invivo. Furthermore, this oligonucleotide demonstrated potent anti-tumoractivity in vivo. Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988)tested a series of tandem 14 nucleobase oligonucleotides, and a 28 and42 nucleobase oligonucleotides comprised of the sequence of two or threeof the tandem oligonucleotides, respectively, for their ability toarrest translation of human DHFR in a rabbit reticulocyte assay. Each ofthe three 14 nucleobase oligonucleotides alone was able to inhibittranslation, albeit at a more modest level than the 28 or 42 nucleobaseoligonucleotides.

In certain embodiments, oligonucleotides are complementary to the targetnucleic acid over the entire length of the oligonucleotide. In certainembodiments, oligonucleotides are 99%, 95%, 90%, 85%, or 80%complementary to the target nucleic acid. In certain embodiments,oligonucleotides are at least 80% complementary to the target nucleicacid over the entire length of the oligonucleotide and comprise a regionthat is 100% or fully complementary to a target nucleic acid. In certainembodiments, the region of full complementarity is from 6 to 20, 10 to18, or 18 to 20 nucleobases in length.

In certain embodiments, oligonucleotides comprise one or more mismatchednucleobases relative to the target nucleic acid. In certain embodiments,antisense activity against the target is reduced by such mismatch, butactivity against a non-target is reduced by a greater amount. Thus, incertain embodiments selectivity of the oligonucleotide is improved. Incertain embodiments, the mismatch is specifically positioned within anoligonucleotide having a gapmer motif. In certain embodiments, themismatch is at position 1, 2, 3, 4, 5, 6, 7, or 8 from the 5′-end of thegap region. In certain embodiments, the mismatch is at position 9, 8, 7,6, 5, 4, 3, 2, 1 from the 3′-end of the gap region. In certainembodiments, the mismatch is at position 1, 2, 3, or 4 from the 5′-endof the wing region. In certain embodiments, the mismatch is at position4, 3, 2, or 1 from the 3′-end of the wing region.

B. SNCA

In certain embodiments, oligomeric compounds comprise or consist of anoligonucleotide comprising a region that is complementary to a targetnucleic acid, wherein the target nucleic acid is SNCA. In certainembodiments, SNCA nucleic acid has the sequence set forth in SEQ ID NO:1 (GENBANK Accession No: NM_000345.3), SEQ ID NO: 2 (GENBANK AccessionNo: NT_016354.20 TRUNC 30800000-30919000), SEQ ID NO: 3 (GENBANKAccession No: JN709863.1), SEQ ID NO: 4 (GENBANK Accession No:BC013293.2), SEQ ID NO: 5 (GENBANK Accession No: NM_001146055.1), andSEQ ID NO: 6 (GENBANK Accession No: HQ830269.1).

In certain embodiments, contacting a cell with an oligomeric compoundcomplementary to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 5, or SEQ ID NO: 6 reduces the amount of SNCA mRNA, and incertain embodiments reduces the amount of alpha-synuclein protein. Incertain embodiments, the oligomeric compound consists of a modifiedoligonucleotide. In certain embodiments, contacting a cell in an animalwith an oligomeric compound complementary to SEQ ID NO: 1, SEQ ID NO: 2,SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6 amelioratesone or more symptom or hallmark of a neurodegenerative disease. Incertain embodiments, the oligomeric compound consists of a modifiedoligonucleotide. In certain embodiments, the symptom or hallmark ismotor dysfunction, aggregation of alpha-synuclein, neurodegeneration,cognitive decline and dementia. In certain embodiments, contacting acell in an animal with an oligonucleotide complementary to SEQ ID NO: 1,SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6results in improved motor function, reduction of alpha-synucleinaggregates, reduced neurodegeneration and/or reduced dementia. Incertain embodiments, the oligomeric compound consists of a modifiedoligonucleotide

C. Certain Target Nucleic Acids in Certain Tissues

In certain embodiments, oligomeric compounds comprise or consist of anoligonucleotide comprising a region that is complementary to a targetnucleic acid, wherein the target nucleic acid is expressed in apharmacologically relevant tissue. In certain embodiments, thepharmacologically relevant tissues are the cells and tissues thatcomprise the central nervous system (CNS). Such cells and tissuesinclude motor cortex, frontal cortex, caudate, amygdala, pons,substantia nigra, putamen, cerebellar peduncle, corpus collosum, dorsalcochlear nucleus (DCN), entorhinal cortex (Ent Cortex), hippocampus,insular cortex, medulla oblongata, central gray matter, pulvinar,occipital cortex, cerebral cortex, temporal cortex, globus pallidus,superior colliculi, and basal forbrain nuclei.

VI. Certain Pharmaceutical Compositions

In certain embodiments, described herein are pharmaceutical compositionscomprising one or more oligomeric compounds. In certain embodiments, theone or more oligomeric compounds each consists of a modifiedoligonucleotide. In certain embodiments, the pharmaceutical compositioncomprises a pharmaceutically acceptable diluent or carrier. In certainembodiments, a pharmaceutical composition comprises or consists of asterile saline solution and one or more oligomeric compound. In certainembodiments, the sterile saline is pharmaceutical grade saline. Incertain embodiments, a pharmaceutical composition comprises or consistsof one or more oligomeric compound and sterile water. In certainembodiments, the sterile water is pharmaceutical grade water. In certainembodiments, a pharmaceutical composition comprises or consists of oneor more oligomeric compound and phosphate-buffered saline (PBS). Incertain embodiments, the sterile PBS is pharmaceutical grade PBS. Incertain embodiments, a pharmaceutical composition comprises or consistsof one or more oligomeric compound and artificial cerebrospinal fluid.In certain embodiments, the artificial cerebrospinal fluid ispharmaceutical grade.

In certain embodiments, a pharmaceutical composition comprises amodified oligonucleotide and artificial cerebrospinal fluid. In certainembodiments, a pharmaceutical composition consists of a modifiedoligonucleotide and artificial cerebrospinal fluid. In certainembodiments, a pharmaceutical composition consists essentially of amodified oligonucleotide and artificial cerebrospinal fluid. In certainembodiments, the artificial cerebrospinal fluid is pharmaceutical grade.

In certain embodiments, pharmaceutical compositions comprise one or moreoligomeric compound and one or more excipients. In certain embodiments,excipients are selected from water, salt solutions, alcohol,polyethylene glycols, gelatin, lactose, amylase, magnesium stearate,talc, silicic acid, viscous paraffin, hydroxymethylcellulose andpolyvinylpyrrolidone.

In certain embodiments, oligomeric compounds may be admixed withpharmaceutically acceptable active and/or inert substances for thepreparation of pharmaceutical compositions or formulations. Compositionsand methods for the formulation of pharmaceutical compositions depend ona number of criteria, including, but not limited to, route ofadministration, extent of disease, or dose to be administered.

In certain embodiments, pharmaceutical compositions comprising anoligomeric compound encompass any pharmaceutically acceptable salts ofthe oligomeric compound, esters of the oligomeric compound, or salts ofsuch esters. In certain embodiments, pharmaceutical compositionscomprising oligomeric compounds comprising one or more oligonucleotide,upon administration to an animal, including a human, are capable ofproviding (directly or indirectly) the biologically active metabolite orresidue thereof. Accordingly, for example, the disclosure is also drawnto pharmaceutically acceptable salts of oligomeric compounds, prodrugs,pharmaceutically acceptable salts of such prodrugs, and otherbioequivalents. Suitable pharmaceutically acceptable salts include, butare not limited to, sodium and potassium salts. In certain embodiments,prodrugs comprise one or more conjugate group attached to anoligonucleotide, wherein the conjugate group is cleaved by endogenousnucleases within the body.

Lipid moieties have been used in nucleic acid therapies in a variety ofmethods. In certain such methods, the nucleic acid, such as anoligomeric compound, is introduced into preformed liposomes orlipoplexes made of mixtures of cationic lipids and neutral lipids. Incertain methods, DNA complexes with mono- or poly-cationic lipids areformed without the presence of a neutral lipid. In certain embodiments,a lipid moiety is selected to increase distribution of a pharmaceuticalagent to a particular cell or tissue. In certain embodiments, a lipidmoiety is selected to increase distribution of a pharmaceutical agent tofat tissue. In certain embodiments, a lipid moiety is selected toincrease distribution of a pharmaceutical agent to muscle tissue.

In certain embodiments, pharmaceutical compositions comprise a deliverysystem. Examples of delivery systems include, but are not limited to,liposomes and emulsions. Certain delivery systems are useful forpreparing certain pharmaceutical compositions including those comprisinghydrophobic compounds. In certain embodiments, certain organic solventssuch as dimethylsulfoxide are used.

In certain embodiments, pharmaceutical compositions comprise one or moretissue-specific delivery molecules designed to deliver the one or morepharmaceutical agents of the present invention to specific tissues orcell types. For example, in certain embodiments, pharmaceuticalcompositions include liposomes coated with a tissue-specific antibody.

In certain embodiments, pharmaceutical compositions comprise aco-solvent system. Certain of such co-solvent systems comprise, forexample, benzyl alcohol, a nonpolar surfactant, a water-miscible organicpolymer, and an aqueous phase. In certain embodiments, such co-solventsystems are used for hydrophobic compounds. A non-limiting example ofsuch a co-solvent system is the VPD co-solvent system, which is asolution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v ofthe nonpolar surfactant Polysorbate 80™ and 65% w/v polyethylene glycol300. The proportions of such co-solvent systems may be variedconsiderably without significantly altering their solubility andtoxicity characteristics. Furthermore, the identity of co-solventcomponents may be varied: for example, other surfactants may be usedinstead of Polysorbate 80™; the fraction size of polyethylene glycol maybe varied; other biocompatible polymers may replace polyethylene glycol,e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides maysubstitute for dextrose.

In certain embodiments, pharmaceutical compositions are prepared fororal administration. In certain embodiments, pharmaceutical compositionsare prepared for buccal administration. In certain embodiments, apharmaceutical composition is prepared for administration by injection(e.g., intravenous, subcutaneous, intramuscular, intrathecal (IT),intracerebroventricular (ICV), etc.). In certain of such embodiments, apharmaceutical composition comprises a carrier and is formulated inaqueous solution, such as water or physiologically compatible bufferssuch as Hanks's solution, Ringer's solution, or physiological salinebuffer. In certain embodiments, other ingredients are included (e.g.,ingredients that aid in solubility or serve as preservatives). Incertain embodiments, injectable suspensions are prepared usingappropriate liquid carriers, suspending agents and the like. Certainpharmaceutical compositions for injection are presented in unit dosageform, e.g., in ampoules or in multi-dose containers. Certainpharmaceutical compositions for injection are suspensions, solutions oremulsions in oily or aqueous vehicles, and may contain formulatoryagents such as suspending, stabilizing and/or dispersing agents. Certainsolvents suitable for use in pharmaceutical compositions for injectioninclude, but are not limited to, lipophilic solvents and fatty oils,such as sesame oil, synthetic fatty acid esters, such as ethyl oleate ortriglycerides, and liposomes. Aqueous injection suspensions may contain.

VII. Certain Compositions

1. Compound No: 763085

In certain embodiments, Compound No: 763085 is characterized as a 5-10-5MOE gapmer, having a sequence of (from 5′ to 3′) CAGACTGTAATCTAGGACCC(incorporated herein as SEQ ID NO: 1887), wherein each of nucleosides1-5 and 16-20 (from 5′ to 3′) comprise a 2′-MOE modification and each ofnucleosides 6-15 are 2′-deoxynucleosides, wherein the internucleosidelinkages between nucleosides 2 to 3, 3 to 4, 4 to 5, 16 to 17, and 17 to18 are phosphodiester internucleoside linkages and the internucleosidelinkages between nucleosides 1 to 2, 5 to 6, 6 to 7, 7 to 8, 8 to 9, 9to 10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15, 15 to 16, 18 to19, and 19 to 20 are phosphorothioate internucleoside linkages, andwherein each cytosine is a 5-methyl cytosine.

In certain embodiments, Compound No: 763085 is characterized by thefollowing chemical notation: mCes Aeo Geo Aeo mCes Tds Gds Tds Ads AdsTds mCds Tds Ads Gds Geo Aeo mCes mCes mCe; wherein,

A=an adenine nucleobase,

mC=a 5-methyl cytosine nucleobase,

G=a guanine nucleobase,

T=a thymine nucleobase,

e=a 2′-MOE modified sugar,

d=a 2′-deoxyribose sugar,

s=a phosphorothioate internucleoside linkage, and

o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No: 763085 is represented by thefollowing chemical structure:

Structure 1. Compound No: 763085

2. Compound No: 763364

In certain embodiments, Compound No: 763364 is characterized as a 5-10-5MOE gapmer, having a sequence of (from 5′ to 3′) ACGACATTTTCTTGCCTCTT(incorporated herein as SEQ ID NO: 2166), wherein each of nucleosides1-5 and 16-20 (from 5′ to 3′) comprise a 2′-MOE modification and each ofnucleosides 6-15 are 2′-deoxynucleosides, wherein the internucleosidelinkages between nucleosides 2 to 3, 3 to 4, 4 to 5, 16 to 17, and 17 to18 are phosphodiester internucleoside linkages and the internucleosidelinkages between nucleosides 1 to 2, 5 to 6, 6 to 7, 7 to 8, 8 to 9, 9to 10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15, 15 to 16, 18 to19, and 19 to 20 are phosphorothioate internucleoside linkages, andwherein each cytosine is a 5-methyl cytosine.

In certain embodiments, Compound No: 763364 is characterized by thefollowing chemical notation: Aes mCeo Geo Aeo mCes Ads Tds Tds Tds TdsmCds Tds Tds Gds mCds mCeo Teo mCes Tes Te; wherein,

A=an adenine nucleobase,

mC=a 5-methyl cytosine nucleobase,

G=a guanine nucleobase,

T=a thymine nucleobase,

e=a 2′-MOE modified sugar,

d=a 2′-deoxyribose sugar,

s=a phosphorothioate internucleoside linkage, and

o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No: 763364 is represented by thefollowing chemical structure:

Structure 2. Compound No: 763364

3. Compound No: 763391

In certain embodiments, Compound No: 763391 is characterized as a 5-10-5MOE gapmer, having a sequence of (from 5′ to 3′) GTTTTCATCAATATCTGCAA(incorporated herein as SEQ ID NO: 2193), wherein each of nucleosides1-5 and 16-20 (from 5′ to 3′) comprise a 2′-MOE modification and each ofnucleosides 6-15 are 2′-deoxynucleosides, wherein the internucleosidelinkages between nucleosides 2 to 3, 3 to 4, 4 to 5, 16 to 17, and 17 to18 are phosphodiester internucleoside linkages and the internucleosidelinkages between nucleosides 1 to 2, 5 to 6, 6 to 7, 7 to 8, 8 to 9, 9to 10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15, 15 to 16, 18 to19, and 19 to 20 are phosphorothioate internucleoside linkages, andwherein each cytosine is a 5-methyl cytosine.

In certain embodiments, Compound No: 763391 is characterized by thefollowing chemical notation: Ges Teo Teo Teo Tes mCds Ads Tds mCds AdsAds Tds Ads Tds mCds Teo Geo mCes Aes Ae; wherein,

-   -   A=an adenine nucleobase,    -   mC=a 5-methyl cytosine nucleobase,    -   G=a guanine nucleobase,    -   T=a thymine nucleobase,    -   e=a 2′-MOE modified sugar,    -   d=a 2′-deoxyribose sugar,    -   s=a phosphorothioate internucleoside linkage, and    -   o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No: 763391 is represented by thefollowing chemical structure:

Structure 3. Compound No: 763391

4. Compound No: 789243

In certain embodiments, Compound No: 789243 is characterized as a 5-10-5MOE gapmer, having a sequence of (from 5′ to 3′) TGAATTCCTTTACACCACAC(incorporated herein as SEQ ID NO: 1639), wherein each of nucleosides1-5 and 16-20 (from 5′ to 3′) comprise a 2′-MOE modification and each ofnucleosides 6-15 are 2′-deoxynucleosides, wherein the internucleosidelinkages between nucleosides 2 to 3 and 17 to 18 are phosphodiesterinternucleoside linkages and the internucleoside linkages betweennucleosides 1 to 2, 3 to 4, 4 to 5, 5 to 6, 6 to 7, 7 to 8, 8 to 9, 9 to10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15, 15 to 16, 16 to17, 18 to 19, and 19 to 20 are phosphorothioate internucleosidelinkages, and wherein each cytosine is a 5-methyl cytosine.

In certain embodiments, Compound No: 789243 is characterized by thefollowing chemical notation: Tes Geo Aes Aes Tes Tds mCds mCds Tds TdsTds Ads mCds Ads mCds mCes Aeo mCes Aes mCe; wherein,

A=an adenine nucleobase,

mC=a 5-methyl cytosine nucleobase,

G=a guanine nucleobase,

T=a thymine nucleobase,

e=a 2′-MOE modified sugar,

d=a 2′-deoxyribose sugar,

s=a phosphorothioate internucleoside linkage, and

o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No: 789243 is represented by thefollowing chemical structure:

Structure 4. Compound No: 789243

5. Compound No: 827599

In certain embodiments, Compound No: 827599 is characterized as a 5-10-5MOE gapmer, having a sequence of (from 5′ to 3′) ACAGATATTTTTGTTCTGCC(incorporated herein as SEQ ID NO: 1703), wherein each of nucleosides1-5 and 16-20 (from 5′ to 3′) comprise a 2′-MOE modification and each ofnucleosides 6-15 are 2′-deoxynucleosides, wherein the internucleosidelinkages between nucleosides 2 to 3, 16 to 17, and 17 to 18 arephosphodiester internucleoside linkages and the internucleoside linkagesbetween nucleosides 1 to 2, 3 to 4, 4 to 5, 5 to 6, 6 to 7, 7 to 8, 8 to9, 9 to 10, 10 to 11, 11 to 12, 12 to 13, 13 to 14, 14 to 15, 15 to 16,18 to 19, and 19 to 20 are phosphorothioate internucleoside linkages,and wherein each cytosine is a 5-methyl cytosine.

In certain embodiments, Compound No: 827599 is characterized by thefollowing chemical notation: Aes mCeo Aes Ges Aes Tds Ads Tds Tds TdsTds Tds Gds Tds Tds mCeo Teo Ges mCes mCe; wherein,

A=an adenine nucleobase,

mC=a 5-methyl cytosine nucleobase,

G=a guanine nucleobase,

T=a thymine nucleobase,

e=a 2′-MOE modified sugar,

d=a 2′-deoxyribose sugar,

s=a phosphorothioate internucleoside linkage, and

o=a phosphodiester internucleoside linkage.

In certain embodiments, Compound No: 827599 is represented by thefollowing chemical structure:

Structure 5. Compound No: 827599

VIII. Certain Comparator Compositions

In certain embodiments, Compound No: 387978, a 5-10-5 MOE gapmer, havinga sequence of (from 5′ to 3′) TCCTTGGCCTTTGAAAGTCC (incorporated hereinas SEQ ID NO: 21), wherein each internucleoside linkage is aphorsphorothioate internucleoside linkage, each cytosine is a 5-methylcytosine, and each of nucleosides 1-5 and 16-20 (from 5′ to 3′) comprisea 2′-MOE modified sugar, which was previously described in WO2012/068405, incorporated herein by reference, is a comparator compound.Compound No. 387978 was selected as a comparator compound because it waspotent in multiple dose studies of reducing human SNCA mRNA withoutovert toxicity in various studies as described in WO 2012/068405. Thus,based on the disclosure of WO 2012/068405, Compound No. 387978 wasdeemed potent with an acceptable tolerability profile.

In certain embodiments, Compound No: 387985, a 5-10-5 MOE gapmer, havinga sequence of (from 5′ to 3′) CCAACATTTGTCACTTGCTC (incorporated hereinas SEQ ID NO: 22), wherein each internucleoside linkage is aphorsphorothioate internucleoside linkage, each cytosine is a 5-methylcytosine, and each of nucleosides 1-5 and 16-20 (from 5′ to 3′) comprisea 2′-MOE modified sugar, which was previously described in WO2012/068405, incorporated herein by reference, is a comparator compound.Compound No. 387985 was selected as a comparator compound because it waspotent in multiple dose studies of reducing human SNCA mRNA withoutovert toxicity in various studies as described in WO 2012/068405. Thus,based on the disclosure of WO 2012/068405, Compound No. 387985 wasdeemed potent with an acceptable tolerability profile.

In certain embodiments, compounds described herein are superior relativeto the compounds described in WO 2012/068405 because they demonstrateone or more improved properties, such as, potency and tolerability.

Compound 763085

For example, as provided in Example 10 (hereinbelow), Compound 763085demonstrated an IC₅₀ of <0.44 μM in SHSH-SYSY cells when tested atconcentrations of 0.44 μM, 1.33 μM, 4.00 μM and 12.00 μM. ComparatorCompound 387985 demonstrated an IC₅₀ of 5.00 μM in the same study.Therefore, Compound 763085 is demonstrably more potent than ComparatorCompound 387985 in this assay.

For example, as provided in Example 11 (hereinbelow), Compound 763085demonstrated an IC₅₀ of 0.47 μM in SHSH-SYSY cells when tested atconcentrations of 0.032 μM, 0.160 μM, 0.800 μM, 4.000 μM and 20.000 μM.Comparator Compound 387985 demonstrated an IC₅₀ of 4.20 μM in the samestudy. Therefore, Compound 763085 is demonstrably more potent thanComparator Compound 387985 in this assay.

For example, as provided in Example 17 (hereinbelow), Compound 763085demonstrated functional observational battery (FOB) scores of 0.8 and1.3 whereas Comparator Compound 387985 demonstrated a FOB score of 6.0in wild-type C57/Bl6 mice after 3 hours when treated with 700 μg ofoligonucleotide by ICV administration. Therefore, Compound 763085 isdemonstrably more tolerable than Comparator Compound 387985 in thisassay.

Compound 763364

For example, as provided in Example 10 (hereinbelow), Compound 763364demonstrated an IC₅₀ of <0.44 μM in SHSH-SYSY cells when tested atconcentrations of 0.44 μM, 1.33 μM, 4.00 μM and 12.00 μM. ComparatorCompound 387985 demonstrated an IC₅₀ of 5.00 μM in the same study.Therefore, Compound 763364 is demonstrably more potent than ComparatorCompound 387985 in this assay.

For example, as provided in Example 11 (hereinbelow), Compound 763364demonstrated an IC₅₀ of 0.86 μM in SHSH-SYSY cells when tested atconcentrations of 0.032 μM, 0.160 μM, 0.800 μM, 4.000 μM and 20.000 μM.Comparator Compound 387985 demonstrated an IC₅₀ of 4.20 μM in the samestudy. Therefore, Compound 763364 is demonstrably more potent thanComparator Compound 387985 in this assay.

Compound 763391

For example, as provided in Example 10 (hereinbelow), Compound 763391demonstrated an IC₅₀ of 0.94 μM and 2.49 μM in SHSH-SYSY cells whentested at concentrations of 0.44 μM, 1.33 μM, 4.00 μM and 12.00 μM.Comparator Compound 387985 demonstrated an IC₅₀ of 5.00 μM in the samestudy. Therefore, Compound 763391 is demonstrably more potent thanComparator Compound 387985 in this assay.

For example, as provided in Example 11 (hereinbelow), Compound 763391demonstrated an IC₅₀ of 1.10 μM in SHSH-SYSY cells when tested atconcentrations of 0.032 μM, 0.160 μM, 0.800 μM, 4.000 μM and 20.000 μM.Comparator Compound 387985 demonstrated an IC₅₀ of 4.20 μM in the samestudy. Therefore, Compound 763391 is demonstrably more potent thanComparator Compound 387985 in this assay.

For example, as provided in Example 17 (hereinbelow), Compound 763391demonstrated a FOB score of 0.0 and 2.3 whereas Comparator Compound387985 demonstrated a FOB score of 6.0 in wild-type C57/Bl6 mice after 3hours when treated with 700 μg of oligonucleotide by ICV administration.Therefore, Compound 763391 is demonstrably more tolerable thanComparator Compound 387985 in this assay.

For example, as provided in Example 18 (hereinbelow), Compound 763391demonstrated FOB scores of 0.0 and 1.3 whereas Comparator Compound387985 demonstrated a FOB score of 3.8 in Sprague Dawley rats after 3hours when treated with 3 mg of oligonucleotide by IT administration.Therefore, Compound 763391 is demonstrably more tolerable thanComparator Compound 387985 in this assay.

Compound 789243

For example, as provided in Example 10 (hereinbelow), Compound 789243demonstrated an IC₅₀ of 2.40 μM in SHSH-SYSY cells when tested atconcentrations of 0.44 μM, 1.33 μM, 4.00 μM and 12.00 μM. ComparatorCompound 387985 demonstrated an IC₅₀ of 5.00 μM in the same study.Therefore, Compound 789243 is demonstrably more potent than ComparatorCompound 387985 in this assay.

For example, as provided in Example 11 (hereinbelow), Compound 789243demonstrated an IC₅₀ of 2.25 μM and 1.90 μM in SHSH-SYSY cells whentested at concentrations of 0.032 μM, 0.160 μM, 0.800 μM, 4.000 μM and20.000 μM. Comparator Compound 387985 demonstrated an IC₅₀ of 4.20 μM inthe same study. Therefore, Compound 789243 is demonstrably more potentthan Comparator Compound 387985 in this assay.

For example, as provided in Example 17 (hereinbelow), Compound 789243demonstrated a FOB score of 0.3 and 0.0 whereas Comparator Compound387985 demonstrated a FOB score of 6.0 in wild-type C57/Bl6 mice after 3hours when treated with 700 μg of oligonucleotide by ICV administration.Therefore, Compound 789243 is demonstrably more tolerable thanComparator Compound 387985 in this assay.

For example, as provided in Example 18 (hereinbelow), Compound 789243demonstrated FOB scores of 1.8 and 1.5 whereas Comparator Compound387985 demonstrated a FOB score of 3.8 in Sprague Dawley rats after 3hours when treated with 3 mg of oligonucleotide by IT administration.Therefore, Compound 789243 is demonstrably more tolerable thanComparator Compound 387985 in this assay.

Compound 827599

For example, as provided in Example 10 (hereinbelow), Compound 827599demonstrated an IC₅₀ of 0.40 μM μM in SHSH-SYSY cells when tested atconcentrations of 0.44 μM, 1.33 μM, 4.00 μM and 12.00 μM. ComparatorCompound 387985 demonstrated an IC₅₀ of 5.00 μM in the same study.Therefore, Compound 827599 is demonstrably more potent than ComparatorCompound 387985 in this assay.

For example, as provided in Example 11 (hereinbelow), Compound 827599demonstrated an IC₅₀ of 0.40 μM in SHSH-SYSY cells when tested atconcentrations of 0.032 μM, 0.160 μM, 0.800 μM, 4.000 μM and 20.000 μM.Comparator Compound 387985 demonstrated an IC₅₀ of 4.20 μM in the samestudy. Therefore, Compound 827599 is demonstrably more potent thanComparator Compound 387985 in this assay.

For example, as provided in Example 17 (hereinbelow), Compound 827599demonstrated a FOB score of 0.0 whereas Comparator Compound 387985demonstrated a FOB score of 6.0 in wild-type C57/Bl6 mice after 3 hourswhen treated with 700 μg of oligonucleotide by ICV administration.Therefore, Compound 827599 is demonstrably more tolerable thanComparator Compound 387985 in this assay.

For example, as provided in Example 18 (hereinbelow), Compound 827599demonstrated a FOB score of 2.0 whereas Comparator Compound 387985demonstrated a FOB score of 3.8 in Sprague Dawley rats after 3 hourswhen treated with 3 mg of oligonucleotide by IT administration.Therefore, Compound 827599 is demonstrably more tolerable thanComparator Compound 387985 in this assay.

IX. Certain Hotspot Regions

1. Nucleobases 50915-50943 of SEQ ID NO: 2

In certain embodiments, nucleobases 50915-50943 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to nucleobases 50915-50943 of SEQ ID NO: 2. In certainembodiments, modified oligonucleotides are 17 or 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are gapmers.In certain embodiments, the gapmers are MOE gapmers or mixed cEt and MOEgapmers. In certain embodiments, the internucleoside linkages of themodified oligonucleotides are phosphorothioate internucleoside linkagesand phosphodiester internucleoside linkages.

The nucleobase sequences of SEQ ID Nos: 243, 1601-1603, and 2188, 2189,2190, 2191, 2192, 2193, 2194, 2195, 2196 and 2197 are complementary tonucleobases 50915-50943 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary tonucleobases 50915-50943 of SEQ ID NO: 2 achieve at least 45% reductionof SNCA RNA in vitro in the standard cell assay.

2. Nucleobases 19630-19656 of SEQ ID NO: 2

In certain embodiments, nucleobases 19630-19656 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to nucleobases 19630-19656 of SEQ ID NO: 2. In certainembodiments, modified oligonucleotides are 17 or 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are gapmers.In certain embodiments, the gapmers are MOE gapmers or mixed cEt and MOEgapmers. In certain embodiments, the nucleosides of the modifiedoligonucleotides are linked by phosphorothioate internucleoside linkagesand phosphodiester internucleoside linkages.

The nucleobase sequences of SEQ ID Nos: 1103, 1700, 1701, 1702, 1703,1704, 1705, 1706 and 1707 are complementary to nucleobases 19630-19656of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary tonucleobases 19630-19656 of SEQ ID NO: 2 achieve at least 48% reductionof SNCA RNA in vitro in the standard cell assay.

3. Nucleobases 28451-28491 of SEQ ID NO: 2

In certain embodiments, nucleobases 28451-28491 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to nucleobases 28451-28491 of SEQ ID NO: 2. In certainembodiments, modified oligonucleotides are 17 or 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are gapmers.In certain embodiments, the gapmers are MOE gapmers or mixed cEt and MOEgapmers. In certain embodiments, the nucleosides of the modifiedoligonucleotides are linked by phosphorothioate internucleoside linkagesand phosphodiester internucleoside linkages.

The nucleobase sequences of SEQ ID Nos: 1168, 1882, 1883, 1884, 1885,1886, 1887, 1888, 1889, 1890, 1891, 1892 and 1893 are complementary tonucleobases 28451-28491 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary tonucleobases 28451-28491 of SEQ ID NO: 2 achieve at least 47% reductionof SNCA RNA in vitro in the standard cell assay.

4. Nucleobases 48712-48760 of SEQ ID NO:2

In certain embodiments, nucleobases 48712-48760 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to nucleobases 48712-48760 of SEQ ID NO: 2. In certainembodiments, modified oligonucleotides are 17 or 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are gapmers.In certain embodiments, the gapmers are MOE gapmers or mixed cEt and MOEgapmers. In certain embodiments, the nucleosides of the modifiedoligonucleotides are linked by phosphorothioate internucleoside linkagesand phosphodiester internucleoside linkages.

The nucleobase sequences of SEQ ID Nos: 471, 1585-1588, and 2157-2166are complementary to nucleobases 48712-48760 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary tonucleobases 48712-48760 of SEQ ID NO: 2 achieve at least 40% reductionof SNCA RNA in vitro in the standard cell assay.

5. Nucleobases 23279-23315 of SEQ ID NO: 2

In certain embodiments, nucleobases 23279-23315 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to nucleobases 23279-23315 of SEQ ID NO: 2. In certainembodiments, modified oligonucleotides are 17 or 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are gapmers.In certain embodiments, the gapmers are MOE gapmers or mixed cEt and MOEgapmers. In certain embodiments, the nucleosides of the modifiedoligonucleotides are linked by phosphorothioate internucleoside linkagesand phosphodiester internucleoside linkages.

The nucleobase sequences of SEQ ID Nos: 164, 1130-1133, and 1797-1810are complementary to nucleobases 23279-23315 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary tonucleobases 23279-23315 of SEQ ID NO: 2 achieve at least 57% reductionof SNCA RNA in vitro in the standard cell assay.

6. Nucleobases 20964-21018 of SEQ ID NO: 2

In certain embodiments, nucleobases 20964-21018 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to nucleobases 20964-21018 of SEQ ID NO: 2. In certainembodiments, modified oligonucleotides are 17 or 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are gapmers.In certain embodiments, the gapmers are MOE gapmers or mixed cEt and MOEgapmers. In certain embodiments, the nucleosides of the modifiedoligonucleotides are linked by phosphorothioate internucleoside linkagesand phosphodiester internucleoside linkages.

The nucleobase sequences of SEQ ID Nos: 391, 468, 1112-1116, and1723-1741 are complementary to nucleobases 20964-21018 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary tonucleobases 20964-21018 of SEQ ID NO: 2 achieve at least 42% reductionof SNCA RNA in vitro in the standard cell assay.

7. Nucleobases 22454-22477 of SEQ ID NO: 2

In certain embodiments, nucleobases 22454-22477 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to nucleobases 22454-22477 of SEQ ID NO: 2. In certainembodiments, modified oligonucleotides are 17 or 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are gapmers.In certain embodiments, the gapmers are MOE gapmers or mixed cEt and MOEgapmers. In certain embodiments, the nucleosides of the modifiedoligonucleotides are linked by phosphorothioate internucleoside linkagesand phosphodiester internucleoside linkages.

The nucleobase sequences of SEQ ID Nos: 88, 1123-1126, and 1778-1782 arecomplementary to nucleobases 22454-22477 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary tonucleobases 22454-22477 of SEQ ID NO: 2 achieve at least 50% reductionof SNCA RNA in vitro in the standard cell assay.

8. Nucleobases 72294-72321 of SEQ ID NO: 2

In certain embodiments, nucleobases 72294-72321 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to nucleobases 72294-72321 of SEQ ID NO: 2. In certainembodiments, modified oligonucleotides are 17 or 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are gapmers.In certain embodiments, the gapmers are MOE gapmers or mixed cEt and MOEgapmers. In certain embodiments, the nucleosides of the modifiedoligonucleotides are linked by phosphorothioate internucleoside linkagesand phosphodiester internucleoside linkages.

The nucleobase sequences of SEQ ID Nos: 1323 and 2345-2353 arecomplementary to nucleobases 72294-72321 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary tonucleobases 72294-72321 of SEQ ID NO: 2 achieve at least 58% reductionof SNCA RNA in vitro in the standard cell assay.

9. Nucleobases 20549-20581 of SEQ ID NO: 2

In certain embodiments, nucleobases 20549-20581 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to nucleobases 20549-20581 of SEQ ID NO: 2. In certainembodiments, modified oligonucleotides are 17 or 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are gapmers.In certain embodiments, the gapmers are MOE gapmers or mixed cEt and MOEgapmers. In certain embodiments, the nucleosides of the modifiedoligonucleotides are linked by phosphorothioate internucleoside linkagesand phosphodiester internucleoside linkages.

The nucleobase sequences of SEQ ID Nos: 314 and 1107-1110 arecomplementary to nucleobases 20549-20581 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary tonucleobases 20549-20581 of SEQ ID NO: 2 achieve at least 58% reductionof SNCA RNA in vitro in the standard cell assay.

10. Nucleobases 27412-27432 of SEQ ID NO: 2

In certain embodiments, nucleobases 27412-27432 of SEQ ID NO: 2 comprisea hotspot region. In certain embodiments, modified oligonucleotides arecomplementary to nucleobases 27412-27432 of SEQ ID NO: 2. In certainembodiments, modified oligonucleotides are 17 or 20 nucleobases inlength. In certain embodiments, modified oligonucleotides are gapmers.In certain embodiments, the gapmers are MOE gapmers or mixed cEt and MOEgapmers. In certain embodiments, the nucleosides of the modifiedoligonucleotides are linked by phosphorothioate internucleoside linkagesand phosphodiester internucleoside linkages.

The nucleobase sequences of SEQ ID Nos: 468, 1113-1114, and 1163 arecomplementary to nucleobases 27412-27432 of SEQ ID NO: 2.

In certain embodiments, modified oligonucleotides complementary tonucleobases 27412-27432 of SEQ ID NO: 2 achieve at least 62% reductionof SNCA RNA in vitro in the standard cell assay.

Nonlimiting Disclosure and Incorporation by Reference

Each of the literature and patent publications listed herein isincorporated by reference in its entirety.

While certain compounds, compositions and methods described herein havebeen described with specificity in accordance with certain embodiments,the following examples serve only to illustrate the compounds describedherein and are not intended to limit the same. Each of the references,GenBank accession numbers, and the like recited in the presentapplication is incorporated herein by reference in its entirety.

Although the sequence listing accompanying this filing identifies eachsequence as either “RNA” or “DNA” as required, in reality, thosesequences may be modified with any combination of chemicalmodifications. One of skill in the art will readily appreciate that suchdesignation as “RNA” or “DNA” to describe modified oligonucleotides is,in certain instances, arbitrary. For example, an oligonucleotidecomprising a nucleoside comprising a 2′-OH sugar moiety and a thyminebase could be described as a DNA having a modified sugar (2′-OH in placeof one 2′-H of DNA) or as an RNA having a modified base (thymine(methylated uracil) in place of a uracil of RNA). Accordingly, nucleicacid sequences provided herein, including, but not limited to those inthe sequence listing, are intended to encompass nucleic acids containingany combination of natural or modified RNA and/or DNA, including, butnot limited to such nucleic acids having modified nucleobases. By way offurther example and without limitation, an oligomeric compound havingthe nucleobase sequence “ATCGATCG” encompasses any oligomeric compoundshaving such nucleobase sequence, whether modified or unmodified,including, but not limited to, such compounds comprising RNA bases, suchas those having sequence “AUCGAUCG” and those having some DNA bases andsome RNA bases such as “AUCGATCG” and oligomeric compounds having othermodified nucleobases, such as “AT^(m)CGAUCG,” wherein ^(m)C indicates acytosine base comprising a methyl group at the 5-position.

Certain compounds described herein (e.g., modified oligonucleotides)have one or more asymmetric center and thus give rise to enantiomers,diastereomers, and other stereoisomeric configurations that may bedefined, in terms of absolute stereochemistry, as (R) or (S), as a orsuch as for sugar anomers, or as (D) or (L), such as for amino acids,etc. Compounds provided herein that are drawn or described as havingcertain stereoisomeric configurations include only the indicatedcompounds. Compounds provided herein that are drawn or described withundefined stereochemistry include all such possible isomers, includingtheir stereorandom and optically pure forms, unless specified otherwise.Likewise, tautomeric forms of the compounds herein are also includedunless otherwise indicated. Unless otherwise indicated, compoundsdescribed herein are intended to include corresponding salt forms.

The compounds described herein include variations in which one or moreatoms are replaced with a non-radioactive isotope or radioactive isotopeof the indicated element. For example, compounds herein that comprisehydrogen atoms encompass all possible deuterium substitutions for eachof the ¹H hydrogen atoms. Isotopic substitutions encompassed by thecompounds herein include but are not limited to: ²H or ³H in place of¹H, ¹³C or ¹⁴C in place of ¹²C, ¹⁵N in place of ¹⁴N, ¹⁷O or ¹⁸O in placeof ¹⁶O, and ³³S, ³⁴S, ³⁵S, or ³⁶S in place of ³²S. In certainembodiments, non-radioactive isotopic substitutions may impart newproperties on the oligomeric compound that are beneficial for use as atherapeutic or research tool. In certain embodiments, radioactiveisotopic substitutions may make the compound suitable for research ordiagnostic purposes such as imaging.

EXAMPLES

The following examples illustrate certain embodiments of the presentdisclosure and are not limiting. Moreover, where specific embodimentsare provided, the inventors have contemplated generic application ofthose specific embodiments. For example, disclosure of anoligonucleotide having a particular motif provides reasonable supportfor additional oligonucleotides having the same or similar motif. And,for example, where a particular high-affinity modification appears at aparticular position, other high-affinity modifications at the sameposition are considered suitable, unless otherwise indicated.

Example 1: Effect of 5-8-4 MOE and cEt Gapmers with MixedInternucleoside Linkages on Human SNCA In Vitro, Single Dose

Modified oligonucleotides complementary to a human SNCA nucleic acidwere designed and tested for their effect on SNCA mRNA in vitro. Themodified oligonucleotides were tested in a series of experiments thathad similar culture conditions.

Cultured SH-SY5Y cells at a density of 20,000 cells per well weretransfected using electroporation with 7,000 nM concentration ofmodified oligonucleotide or no modified oligonucleotide for untreatedcontrols. After approximately 24 hours, RNA was isolated from the cellsand SNCA mRNA levels were measured by quantitative real-time PCR Humanprimer probe set RTS2621 (forward sequence ACGAACCTGAAGCCTAAGAAATATCT,designated herein as SEQ ID NO: 11; reverse sequenceGAGCACTTGTACAGGATGGAACAT, designated herein as SEQ ID NO: 12; probesequence TGCTCCCAAGTTTCTTGAGATCTGCTGACA, designated herein as SEQ ID:13) was used to measure mRNA levels. SNCA mRNA levels were adjustedaccording to total RNA content, as measured by RIBOGREEN®. Results arepresented in the tables below as percent reduction of the amount of SNCAmRNA, relative to untreated control cells (these conditions describe a“Standard Cell Assay”). The modified oligonucleotides marked with anasterisk (*) target the amplicon region of the primer probe set.Additional assays may be used to measure the potency and efficacy ofoligonucleotides targeting the amplicon region. Compound No. 387978,previously disclosed in WO 2012/068405 was also tested and is acomparator oligonucleotide. Compound No. 387978 is a 5-10-5 MOE gapmerwherein each internucleoside linkage is a phosphorothioateinternucleoside linkage and each cytosine residue is a 5-methylcytosine.

The modified oligonucleotides in tables 1-7 are 5-8-4 mixed MOE and cEtgapmers. The gapmers are 17 nucleobases in length, wherein the centralgap segment comprises eight 2′-deoxynucleosides and is flanked by a wingsegment on the 5′ end comprising five 2′-MOE nucleosides and a wingsegment on the 3′ end comprising two cEt nucleosides and two 2′-MOEnucleosides. The sugar motif for the gapmers is (from 5′ to 3′):eeeeeddddddddkkee; wherein ‘d’ represents a 2′-deoxyribose sugar; ‘e’represents a 2′-MOE modified sugar; and ‘k’ represents a cEt modifiedsugar. All cytosine residues throughout each gapmer are 5-methylcytosines. The internucleoside linkages are mixed phosphodiester andphosphorothioate linkages. The internucleoside linkage motif for thegapmers is (from 5′ to 3′): sooosssssssssoss; wherein ‘o’ represents aphosphodiester internucleoside linkage and ‘s’ represents aphosphorothioate internucleoside linkage. “Start Site” indicates the5′-most nucleoside to which the gapmer is complementary in the humannucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside towhich the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in the Tables below iscomplementary to human SNCA nucleic acid sequences SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6, asindicated. ‘N/A’ indicates that the modified oligonucleotide is notcomplementary to that particular nucleic acid with 100% complementarity.A value of 0% reduction indicates that the compound had no effect orincreased mRNA concentrations in the cell. As shown below, modifiedoligonucleotides complementary to human SNCA reduced the amount of humanSNCA mRNA.

TABLE 1Percent reduction of human SNCA mRNA with 5-8-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID % SEQCompound No: 1 No: 1 No: 2 No: 2 Sequence Reduc- ID No start stop startstop (5′ to 3′) tion NO 387978 282 301 4733 4752 TCCTTGGCCTTTGAAAGTCC 6421 709518 9 25 3191 3207 TAGTCCTCCTCCTTCTC 34 30 709524 102 118 32843300 TGCAGCCCGCACGCACC 33 31 709530 232 248 N/A N/A TTACACCACACTGTCGT 4332 709536 240 256 4691 4707 GAATTCCTTTACACCAC 81 33 709542 256 272 47074723 TACATCCATGGCTAATG 53 34 709548 278 294 4729 4745 CCTTTGAAAGTCCTTTC62 35 709554 288 304 4739 4755 CCCTCCTTGGCCTTTGA 54 36 709560 372 388N/A N/A GAGCCTACATAGAGAAC 61 37 709566 385 401 12198 12214CTCCTTGGTTTTGGAGC 22 38 709572 405 421 12218 12234 GCCACACCATGCACCAC 7939 709578 440 456 18007 18023 TTGTCACTTGCTCTTTG 65 40 709584 450 46618017 18033 CCTCCAACATTTGTCAC 23 41 709590 470 486 18037 18053TCACACCCGTCACCACT 58 42 709596 512 528 18079 18095 CAATGCTCCCTGCTCCC 6143 709602 584 600 111119 111135 GAATTCCTTCCTGTGGG 0 44 709608 650 666N/A N/A CTTGATACCCTTCCTCA 0 45 709614* 729 745 113797 113813CTTGTACAGGATGGAAC 1 46 709620 789 805 113857 113873 TTCGAGATACACTGTAA 3447 709626 798 814 113866 113882 ATGGAAGACTTCGAGAT 0 48 709632 866 882113934 113950 TCACTTCAGTGAAAGGG 67 49 709638 892 908 113960 113976CACACAAAGACCCTGCT 0 50 709644 906 922 113974 113990 CACAAAATCCACAGCAC 051 709650 934 950 114002 114018 AATTTGTTTTAACATCG 0 52 709656 956 972114024 114040 TGGTAGTCACTTAGGTG 30 53 709662 1034 1050 114102 114118TCTTATAATATATGATA 0 54 709668 1133 1149 114201 114217 CATAGTTTCATGCTCAC66 55 709674 1213 1229 114281 114297 TTCTCACCATTTATATA 9 56 709680 12771293 114345 114361 TATTATTAAAGTGAGAT 13 57 709686 1327 1343 114395114411 TTTGTCCTTTGTGTCAG 22 58 709692 1410 1426 114478 114494TCCGAGTGTAGGGTTAA 12 59 709698 1476 1492 114544 114560 AATCACAGCCACTTAAG11 60 709704 1590 1606 114658 114674 ACATCAAACAACAGTTC 0 61 709710 17161732 114784 114800 AGGTACAGCATTCACAC 14 62 709716 1744 1760 114812114828 CATGGTCGAATATTATT 25 63 709722 1816 1832 114884 114900AAGGAGGGTGTAGTCAA 40 64 709728 1882 1898 114950 114966 AAGTTAACCACATTCTC5 65 709734 2013 2029 115081 115097 GGTAGTTCCAACGATGT 29 66 709740 20792095 115147 115163 CAACATTTAAAGGAGGC 35 67 709746 2165 2181 115233115249 TTTTCAGCACCCATGGG 2 68 709752 2261 2277 115329 115345GTGACTTTTAGAAATGA 43 69 709758 2327 2343 115395 115411 CTCATGAATACATATAA11 70 709764 2400 2416 115468 115484 TTCTATGGTAACCATCC 37 71 709770 24692485 115537 115553 TAGTGTAAGATGACACA 11 72 709776 2540 2556 115608115624 ACTGTTCAATAACAAAT 33 73 709782 2648 2664 115716 115732TCCTCTATTTCTTAATT 1 74 709788 2714 2730 115782 115798 TAAATTCATGGTCACAA68 75 709794 2783 2799 115851 115867 AAAATTACCGTCAGATA 36 76 709800 28672883 115935 115951 AGGCTTATATGACTTAA 12 77 709806 2933 2949 116001116017 GATTGATCCTCAGGCCA 41 78 709812 2999 3015 116067 116083ACCGTGGAGTCATATGA 0 79 709818 3065 3081 116133 116149 ACACATTAGATTGTTCT16 80 709824 3131 3147 116199 116215 GAAACATGTTTGCATCT 47 81 709836 N/AN/A 3445 3461 GGCGACGCGAGGCTGGG 22 82 709842 N/A N/A 3553 3569ACAATTCCCAAATAATA 6 83 709854 N/A N/A 2097 2113 GACAGCTGTTCCTGGAT 32 84709860 N/A N/A 3957 3973 ACCAAGAGAGCGGGCAG 30 85 709866 N/A N/A 86138629 AAAGAATGCCACTAGGC 23 86 709872 N/A N/A 17660 17676TACAGGTGCAGTTATAT 20 87 709878 N/A N/A 22457 22473 GCCTGTGACCTGTGCTT 8188 709884 N/A N/A 27802 27818 GACATCTCTAACATAAA 56 89 709890 N/A N/A41133 41149 AACAGATTCCAGCAGAG 74 90 709896 N/A N/A 48867 48883GATGGATATTGACTCCT 53 91 709902 N/A N/A 54583 54599 TATATGCATTTTTCAGG 4892 709908 N/A N/A 57557 57573 AGACACTCTTACTTGAG 0 93 709914 N/A N/A71391 71407 TGAAGGACAACTGTGTA 26 94 709922 N/A N/A 75588 75604GACATCTGAAGTGTTCA 59 95 709928 N/A N/A 78911 78927 ATAACCACCACTGAATT 1296 709934 N/A N/A 80751 80767 CCATGCTACATTGCTCA 18 97 709940 N/A N/A83531 83547 GAAAGAACAATGTCATC 75 98 709946 N/A N/A 89651 89667ACAAACCCAAAGAGATT 51 99 N/A N/A 88646 88662 709952 N/A N/A 89681 89697GTCCCCAATCCCCACCC 11 100 N/A N/A 88676 88692 709958 N/A N/A 89722 89738TCCTATAGAGATGAAGT 40 101 N/A N/A 88717 88733 709964 N/A N/A 89731 89747TATCCACTCTCCTATAG 0 102 N/A N/A 88726 88742 709970 N/A N/A 89191 89207TCCTTGAAAACTTCCAT 48 103 709976 N/A N/A 93421 93437 GAGGTCAAATTTTCCAG 30104 709982 N/A N/A 105440 105456 GAGTGACAGTGGTGGGC 28 105

TABLE 2Percent reduction of human SNCA mRNA with 5-8-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID % SEQCompound No: 1 No: 1 No: 2 No: 2 Sequence Reduc- ID No start stop startstop (5′ to 3′) tion NO 387978 282 301 4733 4752 TCCTTGGCCTTTGAAAGTCC 6021 709519 22 38 3204 3220 GTCCTCCTCCTCCTAGT 32 106 709525 168 184 33503366 GGCTTGAAGGCAAGGCG 46 107 709531 233 249 N/A N/A TTTACACCACACTGTCG33 108 709537 242 258 4693 4709 ATGAATTCCTTTACACC 46 109 709543 257 2734708 4724 ATACATCCATGGCTAAT 62 110 709549 280 296 4731 4747GGCCTTTGAAAGTCCTT 72 111 709555 301 317 4752 4768 AGCAGCCACAACTCCCT 75112 709561 377 393 N/A N/A TTTTGGAGCCTACATAG 28 113 709567 387 403 1220012216 CCCTCCTTGGTTTTGGA 46 114 709573 406 422 12219 12235TGCCACACCATGCACCA 66 115 709579 442 458 18009 18025 ATTTGTCACTTGCTCTT 69116 709585 453 469 18020 18036 GCTCCTCCAACATTTGT 57 117 709591 471 48718038 18054 GTCACACCCGTCACCAC 71 118 709597 523 539 18090 18106AGTGGCTGCTGCAATGC 68 119 709603 595 611 111130 111146 CATATCTTCCAGAATTC9 120 709609* 671 687 113739 113755 CTTAGGCTTCAGGTTCG 92 121 709615* 740756 113808 113824 GGAACTGAGCACTTGTA 67 122 709621 790 806 113858 113874CTTCGAGATACACTGTA 31 123 709627 800 816 113868 113884 TGATGGAAGACTTCGAG12 124 709633 877 893 113945 113961 CTACCATGTATTCACTT 53 125 709639 893909 113961 113977 GCACACAAAGACCCTGC 33 126 709645 908 924 113976 113992GCCACAAAATCCACAGC 56 127 709651 944 960 114012 114028 AGGTGTTTTTAATTTGT63 128 709657 967 983 114035 114051 TTAGAAATAAGTGGTAG 28 129 709663 10451061 114113 114129 ACACCTAAAAATCTTAT 26 130 709669 1144 1160 114212114228 ATTTATAGGTGCATAGT 24 131 709675 1217 1233 114285 114301TTAATTCTCACCATTTA 1 132 709681 1279 1295 114347 114363 TTTATTATTAAAGTGAG4 133 709687 1347 1363 114415 114431 GCTATTAATAACTTTAT 6 134 709693 14211437 114489 114505 CTTCAGGGAATTCCGAG 35 135 709699 1487 1503 114555114571 TTTCAATAATTAATCAC 16 136 709705 1628 1644 114696 114712GGCTCAATTAAAAATGT 0 137 709711 1727 1743 114795 114811 TATTGTCAGAAAGGTAC28 138 709717 1749 1765 114817 114833 TTATTCATGGTCGAATA 0 139 7097231827 1843 114895 114911 TATGGCTCTCTAAGGAG 17 140 709729 1893 1909 114961114977 TGAGTTAAACAAAGTTA 6 141 709735 2024 2040 115092 115108AAGGTGACTCTGGTAGT 35 142 709741 2090 2106 115158 115174CATATATTTGGCAACAT 27 143 709747 2177 2193 115245 115261CCATCAAGTTTATTTTC 30 144 709753 2272 2288 115340 115356ACTTTCTACTAGTGACT 16 145 709759 2338 2354 115406 115422ATATCACATTACTCATG 22 146 709765 2414 2430 115482 115498GTAAAAAAGGAAGTTTC 11 147 709771 2480 2496 115548 115564CCATTTCTCTCTAGTGT 16 148 709777 2551 2567 115619 115635TCCTGAAATATACTGTT 5 149 709783 2659 2675 115727 115743 GTCTAGTTCTGTCCTCT34 150 709789 2726 2742 115794 115810 CACATAAATCCTTAAAT 7 151 7097952794 2810 115862 115878 TTCACTGCTCAAAAATT 8 152 709801 2878 2894 115946115962 GCTTCCTGAAAAGGCTT 40 153 709807 2944 2960 116012 116028ACCTAGGACTGGATTGA 1 154 709813 3010 3026 116078 116094 TGGTAAAGCCGACCGTG29 155 709819 3076 3092 116144 116160 AATACCAAACCACACAT 4 156 7098253145 3161 116213 116229 GCCAGAAAGATGAGGAA 13 157 709837 N/A N/A 34563472 CGCTGTGAGCCGGCGAC 0 158 709843 N/A N/A 3586 3602 CCGCCTCTCTCTTTTTT26 159 709855 N/A N/A 2112 2128 CTTTCAGAGCTGGAAGA 3 160 709861 N/A N/A4256 4272 CAGAACTAACTGCTCAC 28 161 709867 N/A N/A 10668 10684AACATCACATGGGCTCA 9 162 709873 N/A N/A 18297 18313 TCTGGGTTAATGCCTGA 62163 709879 N/A N/A 23286 23302 ATTGTTCTCAGAGACCA 72 164 709885 N/A N/A31744 31760 ACAGTAAAGATTTGCAT 29 165 709891 N/A N/A 42838 42854TGATGCCTCTACCTCCA 70 166 709897 N/A N/A 49481 49497 TTGAAATTTTCCAGCTA 69167 N/A N/A 80992 81008 709903 N/A N/A 55047 55063 TATACCTAATATGTTTG 15168 709909 N/A N/A 58992 59008 ATTTCATTAATCTGTGA 63 169 709915 N/A N/A73191 73207 CAGACTTTCTGTGTGGT 77 170 709923 N/A N/A 76780 76796AATTTGGAAGCTAATGT 24 171 709929 N/A N/A 79117 79133 AGTTCCCATGAGACCAG 56172 709935 N/A N/A 81199 81215 TGGCTTGGAGCAAAAGG 42 173 709941 N/A N/A85498 85514 TTATGCAGTGGAACTAA 20 174 709947 N/A N/A 88649 88665CATACAAACCCAAAGAG 0 175 N/A N/A 89654 89670 709953 N/A N/A 88708 88724GATGAAGTTAACTCCCT 62 176 N/A N/A 89713 89729 709959 N/A N/A 88719 88735TCTCCTATAGAGATGAA 0 177 N/A N/A 89724 89740 709965 N/A N/A 88728 88744TCTATCCACTCTCCTAT 35 178 N/A N/A 89733 89749 709971 N/A N/A 89219 89235TCTGTTAACTGAGGTAG 55 179 709977 N/A N/A 93953 93969 GGCTTCTGGCTGACTGA 71180 709983 N/A N/A 106925 106941 GAACATTAAAATTTGCA 25 181

TABLE 3Percent reduction of human SNCA mRNA with 5-8-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 387978  282  301   4733   4752TCCTTGGCCTTTGAAAGTCC 72  21 709520   44   60   3226   3242TGGGCCCCTTCTGGTCG 14 182 709526  179  195   3361   3377GAAAGGCAGAAGGCTTG 27 183 709532  234  250 N/A N/A CTTTACACCACACTGTC 45184 709538  244  260   4695   4711 TAATGAATTCCTTTACA 35 185 709544  258 274   4709   4725 AATACATCCATGGCTAA 75 186 709550  282  298   4733  4749 TTGGCCTTTGAAAGTCC 77 187 709556  312  328   4763   4779GTTTTCTCAGCAGCAGC 72 188 709562  379  395 N/A N/A GGTTTTGGAGCCTACAT 63189 709568  395  411  12208  12224 GCACCACTCCCTCCTTG 55 190 709574  408 424  12221  12237 GTTGCCACACCATGCAC 49 191 709580  444  460  18011 18027 ACATTTGTCACTTGCTC 77 192 709586  464  480  18031  18047CCGTCACCACTGCTCCT 83 193 709592  473  489  18040  18056CTGTCACACCCGTCACC 78 194 709598  534  550  18101  18117TTGACAAAGCCAGTGGC 31 195 709604  606  622 111141 111157GGATCCACAGGCATATC 29 196 709610*  682  698 113750 113766CAAAGATATTTCTTAGG 23 197 709616*  751  767 113819 113835CTGGGCACATTGGAACT  4 198 709622  792  808 113860 113876GACTTCGAGATACACTG 57 199 709628  811  827 113879 113895TCAATCACTGCTGATGG 45 200 709634  887  903 113955 113971AAAGACCCTGCTACCAT 52 201 709640  895  911 113963 113979CAGCACACAAAGACCCT 65 202 709646  909  925 113977 113993AGCCACAAAATCCACAG 56 203 709652  945  961 114013 114029TAGGTGTTTTTAATTTG 52 204 709658  978  994 114046 114062ATAGTGAGGATTTAGAA 19 205 709664 1056 1072 114124 114140ATCATTAAAAGACACCT 37 206 709670 1172 1188 114240 114256CGCAAAATGGTAAAATT 25 207 709676 1226 1242 114294 114310CGTTTTATTTTAATTCT 27 208 709682 1294 1310 114362 114378CTTATAAGCATGATTTT 10 209 709688 1359 1375 114427 114443CTTCTTCAAATGGCTAT 37 210 709694 1432 1448 114500 114516TGGCAGTGTTGCTTCAG 63 211 709700 1520 1536 114588 114604CTACAATAGTAGTTGGG 24 212 709706 1639 1655 114707 114723TGTTAATAAAAGGCTCA 46 213 709712 1730 1746 114798 114814ATTTATTGTCAGAAAGG 62 214 709718 1772 1788 114840 114856GGGAACCCACTTTTTTT 17 215 709724 1838 1854 114906 114922CTAATGTGTCTTATGGC 39 216 709730 1904 1920 114972 114988GTGAGGAATGCTGAGTT 25 217 709736 2035 2051 115103 115119TGATCTCCTTTAAGGTG 10 218 709742 2107 2123 115175 115191GGAAAAATCCTAGAATT  6 219 709748 2188 2204 115256 115272AGAGTTTTTCACCATCA 37 220 709754 2283 2299 115351 115367CTTGAAATTATACTTTC 37 221 709760 2349 2365 115417 115433GCGCCCAATATATATCA 30 222 709766 2425 2441 115493 115509TCTTCAATTAGGTAAAA 17 223 709772 2491 2507 115559 115575CAAGAAACTTACCATTT 11 224 709778 2562 2578 115630 115646CTTTCTAACCTTCCTGA 21 225 709784 2670 2686 115738 115754CACTGCTATCAGTCTAG 39 226 709790 2737 2753 115805 115821GAATTTGTATCCACATA 53 227 709796 2806 2822 115874 115890TATATAAAGTAATTCAC 12 228 709802 2889 2905 115957 115973ATATGAGACAAGCTTCC 18 229 709808 2955 2971 116023 116039CTGCAAAATAAACCTAG 41 230 709814 3021 3037 116089 116105CTGAACTGTTTTGGTAA 30 231 709820 3087 3103 116155 116171ACCCCACTTGGAATACC 37 232 709826 3156 3172 116224 116240ATACTGGATAAGCCAGA 22 233 709832 N/A N/A  18121  18137 CCTTGCCCAACTGGTCC42 234 709838 N/A N/A   3467   3483 CCAGAGGAGGCCGCTGT 15 235 709844 N/AN/A   3597   3613 CCGACTCCTCCCCGCCT 24 236 709862 N/A N/A   7047   7063TCTTTCCACTCTATCAG 19 237 709868 N/A N/A  10846  10862 ACTGCATATTTAGAGTC13 238 709874 N/A N/A  18424  18440 ACATGAAAGCCCTCATT 37 239 709880 N/AN/A  25537  25553 ATGAATTGCCACTATAA 56 240 709886 N/A N/A  32984  33000TGGATAAAAGAAGTTAC 61 241 709892 N/A N/A  43821  43837 TACTTCTCTGGACCTCT74 242 709898 N/A N/A  50921  50937 TTTCATCAATATCTGCA 90 243 709904 N/AN/A  55614  55630 AATTTAACCTTAAAGTA 20 244 709910 N/A N/A  59199  59215GTAGAGGCCCAATAAGT 37 245 709916 N/A N/A  74075  74091 AGTTATTGCTATCAAGA57 246 709924 N/A N/A  77666  77682 GACTCTAGAAAAGCTCT 70 247 709930 N/AN/A  79403  79419 CTTTTTCACTTGTCTCA 48 248 709936 N/A N/A  81475  81491AGAGCTGTTTGAAGTGA 71 249 709942 N/A N/A  85512  85528 ACCTATGTTGAAACTTA48 250 709948 N/A N/A  88651  88667 GACATACAAACCCAAAG 46 251 N/A N/A 89656  89672 709954 N/A N/A  88711  88727 AGAGATGAAGTTAACTC 61 252 N/AN/A  89716  89732 709960 N/A N/A  88720  88736 CTCTCCTATAGAGATGA 44 253N/A N/A  89725  89741 709966 N/A N/A  88757  88773 CCCTTTTCAAGAGCTTT 82254 N/A N/A  89762  89778 709972 N/A N/A  89254  89270 TAAGCTCATATTTATAG23 255 709978 N/A N/A  94005  94021 TAAAAGATCATGAGGGC 47 256 709984 N/AN/A 107602 107618 GAAACATGAGTTTAATA  9 257

TABLE 4Percent reduction of human SNCA mRNA with 5-8-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 387978  282  301   4733   4752TCCTTGGCCTTTGAAAGTCC 71  21 709521   55   71   3237   3253GCCCCCTCTCTTGGGCC  8 258 709527  190  206   3372   3388TCACGAGGGTGGAAAGG  0 259 709533  236  252   4687   4703TCCTTTACACCACACTG 63 260 709539  252  268   4703   4719TCCATGGCTAATGAATT 24 261 709545  260  276   4711   4727TGAATACATCCATGGCT 46 262 709551  283  299   4734   4750CTTGGCCTTTGAAAGTC 72 263 709557  325  341   4776   4792CACACCCTGTTTGGTTT 45 264 709563  381  397  12194  12210TTGGTTTTGGAGCCTAC 69 265 709569  400  416  12213  12229ACCATGCACCACTCCCT 41 266 709575  410  426  12223  12239CTGTTGCCACACCATGC 45 267 709581  445  461  18012  18028AACATTTGTCACTTGCT 65 268 709587  465  481  18032  18048CCCGTCACCACTGCTCC 24 269 709593  475  491  18042  18058TGCTGTCACACCCGTCA 55 270 709599  545  561  18112  18128ACTGGTCCTTTTTGACA 15 271 709605  617  633 111152 111168CCTCATTGTCAGGATCC 42 272 709611  693  709 113761 113777GAAACTGGGAGCAAAGA 23 273 709617  762  778 113830 113846AAATGTCATGACTGGGC 37 274 709623  793  809 113861 113877AGACTTCGAGATACACT 36 275 709629  822  838 113890 113906GTACAGATACTTCAATC 34 276 709635  888  904 113956 113972CAAAGACCCTGCTACCA  8 277 709641  897  913 113965 113981CACAGCACACAAAGACC 22 278 709647  910  926 113978 113994AAGCCACAAAATCCACA 41 279 709653  947  963 114015 114031CTTAGGTGTTTTTAATT 19 280 709659 1001 1017 114069 114085TTCTGAACAACAGCAAC 41 281 709665 1067 1083 114135 114151TCTTAGACAGTATCATT 38 282 709671 1183 1199 114251 114267ATAAAACACATCGCAAA 13 283 709677 1241 1257 114309 114325TTTGCAATGAGATAACG  8 284 709683 1295 1311 114363 114379GCTTATAAGCATGATTT 22 285 709689 1370 1386 114438 114454TAAAATTCCTCCTTCTT  4 286 709695 1443 1459 114511 114527AAACACACTTCTGGCAG 19 287 709701 1531 1547 114599 114615AATAGACCACTCTACAA 16 288 709707 1660 1676 114728 114744CGAGACAAAAATAACAA  0 289 709713 1735 1751 114803 114819ATATTATTTATTGTCAG  0 290 709719 1783 1799 114851 114867GCTTAGTTCCCGGGAAC  5 291 709725 1849 1865 114917 114933GCTAATATGTGCTAATG 29 292 709731 1930 1946 114998 115014GAATTTCTGATGATTAA  9 293 709737 2046 2062 115114 115130GTCTAGAGAATTGATCT 23 294 709743 2118 2134 115186 115202ACCTTTCCTAAGGAAAA  0 295 709749 2200 2216 115268 115284ATTAATTTATACAGAGT  5 296 709755 2294 2310 115362 115378GAATATTCTGTCTTGAA 30 297 709761 2362 2378 115430 115446TCCTTCCTCACCAGCGC 31 298 709767 2436 2452 115504 115520GTAGTAGTCTCTCTTCA 26 299 709773 2507 2523 115575 115591CATAACTTAAATAAAAC  0 300 709779 2573 2589 115641 115657CCTAACCGCCACTTTCT 19 301 709785 2681 2697 115749 115765TTGTTCTAGGTCACTGC 27 302 709791 2749 2765 115817 115833CACTTTAAAGGAGAATT  0 303 709797 2827 2843 115895 115911GTCCCAAATAAACTATT  0 304 709803 2900 2916 115968 115984CTCGGGAGTGAATATGA 12 305 709809 2966 2982 116034 116050AGAATGTAAGTCTGCAA 24 306 709815 3032 3048 116100 116116CAAAGTGCACTCTGAAC 23 307 709821 3098 3114 116166 116182TTCTGAAAAAGACCCCA  0 308 709827 3167 3183 116235 116251CAAATAGCTACATACTG  2 309 709839 N/A N/A   3495   3511 CGGAGGCGGCACCCGGG 8 310 709845 N/A N/A   3608   3624 TCTCCACAACTCCGACT 18 311 709863 N/AN/A   7156   7172 TAATCAGGGAAGTGATG  0 312 709869 N/A N/A  14963  14979CTTCAGAAAATCTCCAG 33 313 709875 N/A N/A  20562  20578 CACAACTATGCTGCAAT58 314 709881 N/A N/A  25804  25820 ATCATCCAGTAGAGTGA 66 315 709887 N/AN/A  33591  33607 GAGAACACTTAAGTGAA 44 316 709893 N/A N/A  46160  46176ATTTCCATGAAGCCAAG 88 317 N/A N/A  53645  53661 709899 N/A N/A  51477 51493 CTAGAGACCACCTGAGA 27 318 709905 N/A N/A  56363  56379CTAATGAACAGAGAAAG  2 319 709911 N/A N/A  68799  68815 CCAAAGTAAGAGGAGAT37 320 709917 N/A N/A  74219  74235 TGTTGCTAAGCACAAAC 44 321 709925 N/AN/A  78068  78084 TCAAGGTGCCATATCTG 51 322 709931 N/A N/A  80286  80302AAAGAAAGGCAGTGTTG  1 323 709937 N/A N/A  82460  82476 CCAATATGGATTCAGCA46 324 709943 N/A N/A  86783  86799 CAATTATTAGCAGTTAC 55 325 709949 N/AN/A  88653  88669 TTGACATACAAACCCAA 60 326 N/A N/A  89658  89674 709955N/A N/A  88713  88729 ATAGAGATGAAGTTAAC 49 327 N/A N/A  89718  89734709961 N/A N/A  88722  88738 CACTCTCCTATAGAGAT  0 328 N/A N/A  89727 89743 709967 N/A N/A  88759  88775 TTCCCTTTTCAAGAGCT 77 329 N/A N/A 89764  89780 709973 N/A N/A  91417  91433 AGAAGGAATGCACAATA 37 330709979 N/A N/A  94055  94071 GCCATAATTCAAGTCAG 63 331 709985 N/A N/A108049 108065 ACTGACAACTTACAGCA 30 332

TABLE 5Percent reduction of human SNCA mRNA with 5-8-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 387978  282  301   4733   4752TCCTTGGCCTTTGAAAGTCC 58  21 709522   66   82   3248   3264CTCGGTCGCTCGCCCCC 36 333 709528  201  217   3383   3399CAGTTCTCCGCTCACGA 26 334 709534  237  253   4688   4704TTCCTTTACACCACACT 60 335 709540  254  270   4705   4721CATCCATGGCTAATGAA 47 336 709546  262  278   4713   4729CATGAATACATCCATGG 31 337 709552  284  300   4735   4751CCTTGGCCTTTGAAAGT 41 338 709558  336  352   4787   4803GCTGCTTCTGCCACACC 61 339 709564  382  398  12195  12211CTTGGTTTTGGAGCCTA 63 340 709570  402  418  12215  12231ACACCATGCACCACTCC 56 341 709576  418  434  N/A N/A CTCAGCCACTGTTGCCA 57342 709582  446  462  18013  18029 CAACATTTGTCACTTGC 72 343 709588  467 483  18034  18050 CACCCGTCACCACTGCT 56 344 709594  486  502  18053 18069 TTCTGGGCTACTGCTGT 11 345 709600  556  572 N/A N/AATTCTTGCCCAACTGGT  7 346 709606  628  644 111163 111179CATTTCATAAGCCTCAT 12 347 709612*  704  720 113772 113788GCAGATCTCAAGAAACT 45 348 709618  778  794 113846 113862CTGTAAAAACTTTGAGA 27 349 709624  794  810 113862 113878AAGACTTCGAGATACAC 43 350 709630  844  860 113912 113928CCGAAATGCTGAGTGGG 29 351 709636  889  905 113957 113973ACAAAGACCCTGCTACC 14 352 709642  899  915 113967 113983TCCACAGCACACAAAGA 30 353 709648  912  928 113980 113996TGAAGCCACAAAATCCA 28 354 709654  949  965 114017 114033CACTTAGGTGTTTTTAA 26 355 709660 1012 1028 114080 114096TCACTAACAACTTCTGA 34 356 709666 1091 1107 114159 114175ACAAATTTCACAATACG 32 357 709672 1198 1214 114266 114282TACAAACACAAGTGAAT 14 358 709678 1266 1282 114334 114350TGAGATGGGATAAAAAT  9 359 709684 1305 1321 114373 114389AATTCATGTTGCTTATA  0 360 709690 1383 1399 114451 114467TCTCTACCTCTTCTAAA  0 361 709696 1454 1470 114522 114538AGTGCATACCAAAACAC  9 362 709702 1549 1565 114617 114633GACAGGATTGAAGGGAG 26 363 709708 1671 1687 114739 114755AAAAATTATTTCGAGAC  0 364 709714 1738 1754 114806 114822CGAATATTATTTATTGT  0 365 709720 1794 1810 114862 114878TCTTCTACACTGCTTAG 14 366 709726 1860 1876 114928 114944GCCTTGAATGTGCTAAT 38 367 709732 1991 2007 115059 115075GGCATTTCCTGTAAAAA 21 368 709738 2057 2073 115125 115141AATTTTTATCAGTCTAG 45 369 709744 2137 2153 115205 115221TCTTCCCTGAAAGAGAA  6 370 709750 2239 2255 115307 115323CCCCAGAATAATTAAAA  1 371 709756 2305 2321 115373 115389TAGCATGTCTAGAATAT 23 372 709762 2378 2394 115446 115462GTCACTCATTCCTCCTT 24 373 709768 2447 2463 115515 115531CTTAGCACTCTGTAGTA  9 374 709774 2518 2534 115586 115602CCTTGCTTAAACATAAC 16 375 709780 2596 2612 115664 115680CTTTAGGTAGATTTAAA  0 376 709786 2692 2708 115760 115776CTAATCTCAAATTGTTC 12 377 709792 2761 2777 115829 115845TAAGGGAAGAAACACTT 11 378 709798 2838 2854 115906 115922TTAAGTGTTTGGTCCCA 53 379 709804 2911 2927 115979 115995CAGGTGAATGTCTCGGG 29 380 709810 2977 2993 116045 116061AATAACTTGGGAGAATG  6 381 709816 3043 3059 116111 116127CAATTGTGTGCCAAAGT 15 382 709822 3109 3125 116177 116193TAGTGCAGAGATTCTGA 38 383 709828 3174 3190 116242 116258TATGTCACAAATAGCTA  3 384 709834 N/A N/A   3415   3431 AACCCGCTAACCTGTCG16 385 709840 N/A N/A   3506   3522 CACAGGAAGGGCGGAGG 10 386 709846 N/AN/A   3619   3635 GTCCCTCTGCTTCTCCA  4 387 709858 N/A N/A   2166   2182CATACACACGCGAACTT  4 388 709864 N/A N/A   7240   7256 TCAATTATTCATATGTC18 389 709870 N/A N/A  15701  15717 CTGCACAGTAAAATGTA  8 390 709876 N/AN/A  20986  21002 AGTGTGAGCAAACATTC 50 391 709876 N/A N/A  27411  27427AGTGTGAGCAAACATTC 50 391 709882 N/A N/A  25926  25942 AATTGAATACATTGTCT63 392 709888 N/A N/A  39106  39122 TCCTAAAGTATTGCACT 31 393 709894 N/AN/A  48228  48244 CCTGGTCATGACTCTGA 62 394 709900 N/A N/A  52420  52436GATCAAATGTATAGAGA 62 395 709906 N/A N/A  56773  56789 AGAGGCAGGGCTAGACA13 396 709912 N/A N/A  68801  68817 TGCCAAAGTAAGAGGAG 56 397 709919 N/AN/A  74295  74311 ATAGAACTCTGTAGTCA 72 398 709926 N/A N/A  78080  78096CAAATGAACTTCTCAAG  7 399 709932 N/A N/A  80397  80413 AAATTACACTGTTGAAT32 400 709938 N/A N/A  82770  82786 GGCAAAGGGCTCTGGTG 34 401 709944 N/AN/A  87946  87962 GTAAGTTGTGACCATGC 76 402 709950 N/A N/A  88655  88671ACTTGACATACAAACCC 45 403 709950 N/A N/A  89660  89676 ACTTGACATACAAACCC45 403 709956 N/A N/A  88714  88730 TATAGAGATGAAGTTAA 26 404 N/A N/A 89719  89735 709962 N/A N/A  88723  88739 CCACTCTCCTATAGAGA 18 405 N/AN/A  89728  89744 709968 N/A N/A  88761  88777 ATTTCCCTTTTCAAGAG 19 406N/A N/A  89766  89782 19 406 709974 N/A N/A  92159  92175TAACTCCATTTAATTGT 17 407 709980 N/A N/A  99285  99301 ACAGTACACTATTTGTT25 408 709986 N/A N/A 109588 109604 ACCACCCCAAACTACCT  0 409

TABLE 6Percent reduction of human SNCA mRNA with 5-8-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 387978  282  301   4733   4752TCCTTGGCCTTTGAAAGTCC 64  21 709523   91  107   3273   3289CGCACCTCACTTCCGCG 20 410 709529  212  228   3394   3410ATGGCCACTCCCAGTTC 21 411 709535  238  254   4689   4705ATTCCTTTACACCACAC 84 412 709541  255  271   4706   4722ACATCCATGGCTAATGA 43 413 709547  266  282   4717   4733CTTTCATGAATACATCC 81 414 709553  286  302   4737   4753CTCCTTGGCCTTTGAAA 31 415 709559  361  377   4812   4828GAGAACACCCTCTTTTG 15 416 709565  383  399  12196  12212CCTTGGTTTTGGAGCCT 63 417 709571  404  420  12217  12233CCACACCATGCACCACT 63 418 709577  431  447  17998  18014GCTCTTTGGTCTTCTCA 45 419 709583  448  464  18015  18031TCCAACATTTGTCACTT 22 420 709589  469  485  18036  18052CACACCCGTCACCACTG 48 421 709595  501  517  18068  18084GCTCCCTCCACTGTCTT 28 422 709601  567  583 N/A N/A GCTCCTTCTTCATTCTT  5423 709607  639  655 N/A N/A TCCTCAGAAGGCATTTC  0 424 709613*  715  731113783 113799 AACATCTGTCAGCAGAT 56 425 709619  788  804 113856 113872TCGAGATACACTGTAAA 18 426 709625  796  812 113864 113880GGAAGACTTCGAGATAC 64 427 709631  855  871 113923 113939AAAGGGAAGCACCGAAA 25 428 709637  891  907 113959 113975ACACAAAGACCCTGCTA 12 429 709643  904  920 113972 113988CAAAATCCACAGCACAC 50 430 709649  914  930 113982 113998ATTGAAGCCACAAAATC 13 431 709655  952  968 114020 114036AGTCACTTAGGTGTTTT 32 432 709661 1023 1039 114091 114107ATGATAGCAAATCACTA 22 433 709667 1122 1138 114190 114206GCTCACATATTTTTAAG 18 434 709673 1209 1225 114277 114293CACCATTTATATACAAA 20 435 709679 1274 1290 114342 114358TATTAAAGTGAGATGGG 28 436 709685 1316 1332 114384 114400TGTCAGTTCTTAATTCA 17 437 709691 1399 1415 114467 114483GGTTAATGTTCCATTTT 29 438 709697 1465 1481 114533 114549CTTAAGGAACCAGTGCA 30 439 709703 1579 1595 114647 114663CAGTTCCCCAAAATACG  0 440 709709 1705 1721 114773 114789TCACACCAATATCAGAC 36 441 709715 1740 1756 114808 114824GTCGAATATTATTTATT 11 442 709721 1805 1821 114873 114889AGTCAAAATCATCTTCT 17 443 709727 1871 1887 114939 114955ATTCTCTCAGAGCCTTG 72 444 709733 2002 2018 115070 115086CGATGTTTAAAGGCATT  0 445 709739 2068 2084 115136 115152GGAGGCCATGAAATTTT 39 446 709745 2148 2164 115216 115232GAGTTAATAGATCTTCC 36 447 709751 2250 2266 115318 115334AAATGACTATGCCCCAG 44 448 709757 2316 2332 115384 115400ATATAAACTGCTAGCAT 23 449 709763 2389 2405 115457 115473CCATCCTTATAGTCACT 48 450 709769 2458 2474 115526 115542GACACATGCAGCTTAGC 42 451 709775 2529 2545 115597 115613ACAAATCCTTTCCTTGC 12 452 709781 2631 2647 115699 115715TAATACCAATACTTTTA 21 453 709787 2703 2719 115771 115787TCACAACTTTCCTAATC  0 454 709793 2772 2788 115840 115856CAGATAAATATTAAGGG  0 455 709799 2856 2872 115924 115940ACTTAAAGAACTTTTTG 11 456 709805 2922 2938 115990 116006AGGCCACTTGGCAGGTG 16 457 709811 2988 3004 116056 116072ATATGAGGCTGAATAAC 16 458 709817 3054 3070 116122 116138TGTTCTGTTCCCAATTG 48 459 709823 3120 3136 116188 116204GCATCTCACACTAGTGC 30 460 709829 3180 3196 116248 116264ATTTATTATGTCACAAA  0 461 709835 N/A N/A   3426   3442 AGTGGGAGGCAAACCCG 8 462 709841 N/A N/A   3517   3533 GAAAAGGAGCGCACAGG 32 463 709853 N/AN/A   2086   2102 CTGGATCACACCAGAAT 16 464 709859 N/A N/A   2500   2516CTATCACCATTTTCCTT 13 465 N/A N/A 112970 112986 709865 N/A N/A   7406  7422 AGCCATAAGTGAAATTA 48 466 709871 N/A N/A  15993  16009AGTTCGATTTAAATGCC 27 467 709877 N/A N/A  20988  21004 ACAGTGTGAGCAAACAT62 468 N/A N/A  27413  27429 709883 N/A N/A  26205  26221CCCTCTTTGTGTTATAC 74 469 709889 N/A N/A  40203  40219 GAAAGTTTTTATGGAGA22 470 709895 N/A N/A  48716  48732 TGTATTTTGGATGCTTC 85 471 709901 N/AN/A  52979  52995 GAAGTGACTATGTCTTC 46 472 709907 N/A N/A  57491  57507GCCAAATGAATGGGCCA 59 473 709913 N/A N/A  68942  68958 ATCAAAAGGAACATCAA35 474 709921 N/A N/A  75328  75344 TATTCTTCTCCTCCATG 34 475 709927 N/AN/A  78404  78420 AATGTTGGCAAGCTTGA 48 476 709933 N/A N/A  80489  80505ACTCACACTGCCTAGCT 36 477 709939 N/A N/A  83333  83349 CCTATATATTCAAGATG22 478 709945 N/A N/A  88047  88063 AGAAGCTATCAAGACAT 60 479 709951 N/AN/A  88657  88673 CCACTTGACATACAAAC 24 480 N/A N/A  89662  89678 709957N/A N/A  88715  88731 CTATAGAGATGAAGTTA 39 481 709957 N/A N/A  89720 89736 709963 N/A N/A  88725  88741 ATCCACTCTCCTATAGA  6 482 709963 N/AN/A  89730  89746 709969 N/A N/A  89098  89114 AATAGGAGTTCAATGAA 33 483709975 N/A N/A  93354  93370 GTTAGATAATTATTGAG 20 484 709981 N/A N/A100015 100031 CTTCAAACCTTTTGACC 15 485 709987 N/A N/A 110359 110375CCTATTTATGGTATAAT  0 486

TABLE 7Percent reduction of human SNCA mRNA with 5-8-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ SEQ SEQ SEQ SEQ SEQ SEQ SEQ IDID ID ID ID ID ID ID Compound No: 3 No: 3 No: 4 No: 4 No: 5 No: 5 No: 6No: 6 % SEQ No start stop start stop start stop start stopSequence (5′ to 3′) Reduction ID NO 709830 369 385 N/A N/A N/A N/A N/AN/A ACTACATAGAGAACACC 60 487 709831 380 396 N/A N/A N/A N/A N/A N/ATCTTCTCAGCCACTACA 23 488 709833 523 539 N/A N/A N/A N/A N/A N/ATTGATACCCTTCCTTGC  0 489 709847 N/A N/A 388 404 N/A N/A N/A N/AACCACACTGAGTCCCTC  0 490 709848 N/A N/A 389 405 N/A N/A N/A N/ACACCACACTGAGTCCCT 22 491 709849 N/A N/A 390 406 N/A N/A N/A N/AACACCACACTGAGTCCC 11 492 709850 N/A N/A 392 408 N/A N/A N/A N/ATTACACCACACTGAGTC 26 493 709851 N/A N/A 393 409 N/A N/A N/A N/ATTTACACCACACTGAGT 23 494 709852 N/A N/A 394 410 N/A N/A N/A N/ACTTTACACCACACTGAG 24 495 709856 N/A N/A N/A N/A 38 54 N/A N/ATACACCACACTCTTTCA 22 496 709857 N/A N/A N/A N/A N/A N/A 89 105CCACACTCACTTCCGCG 15 497

Example 2: Effect of 4-9-4 MOE and cEt Gapmers with MixedInternucleoside Linkages on Human SNCA In Vitro, Single Dose

Modified oligonucleotides complementary to a human SNCA nucleic acidwere designed and tested as described in Example 1 for their effect onSNCA mRNA in vitro. The modified oligonucleotides were tested in aseries of experiments that had similar culture conditions.

The modified oligonucleotides marked with an asterisk (*) target theamplicon region of the primer probe set. Additional assays may be usedto measure the potency and efficacy of oligonucleotides targeting theamplicon region. Compound No. 387978, previously disclosed in WO2012/068405 was also tested and is a comparator oligonucleotide.Compound No. 387978 is a 5-10-5 MOE gapmer wherein each internucleosidelinkage is a phosphorothioate internucleoside linkage and each cytosineresidue is a 5-methyl cytosine.

The modified oligonucleotides in tables 7-13 are 4-9-4 MOE and cEtgapmers. The gapmers are 17 nucleobases in length, wherein the centralgap segment comprises nine 2′-deoxynucleosides and is flanked by wingsegments on both the 5′ end on the 3′ end comprising two 2′-MOEnucleosides and two cEt nucleosides. The sugar motif for the gapmers is(from 5′ to 3′): eekkdddddddddkkee; wherein ‘d’ represents a2′-deoxyribose sugar; ‘e’ represents a 2′-MOE modified sugar; and ‘k’represents a cEt modified sugar. All cytosine residues throughout eachgapmer are 5-methyl cytosines. The internucleoside linkages are mixedphosphodiester and phosphorothioate linkages. The internucleosidelinkage motif for the gapmers is (from 5′ to 3′): sooosssssssssoss;wherein ‘o’ represents a phosphodiester internucleoside linkage and ‘s’represents a phosphorothioate internucleoside linkage. “Start Site”indicates the 5′-most nucleoside to which the gapmer is complementary inthe human nucleic acid sequence. “Stop Site” indicates the 3′-mostnucleoside to which the gapmer is complementary in the human nucleicacid sequence.

Each modified oligonucleotide listed in the Tables below iscomplementary to human SNCA nucleic acid sequences SEQ ID NO: 1 or SEQID NO: 2, as indicated. ‘N/A’ indicates that the modifiedoligonucleotide is not complementary to that particular nucleic acidwith 100% complementarity. A value of 0% reduction indicates that thecompound had no effect or increased mRNA concentrations in the cell. Asshown below, modified oligonucleotides complementary to human SNCAreduced the amount of human SNCA mRNA.

TABLE 8Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 387978 282 301 4733 4752TCCTTGGCCTTTGAAAGTCC  9  21 740410 240 256 4691 4707 GAATTCCTTTACACCAC92  33 740316 N/A N/A 3416 3432 AAACCCGCTAACCTGTC 16 498 740317 N/A N/A3419 3435 GGCAAACCCGCTAACCT 47 499 740318 N/A N/A 3422 3438GGAGGCAAACCCGCTAA 13 500 740319 N/A N/A 3425 3441 GTGGGAGGCAAACCCGC  2501 740320 N/A N/A 3428 3444 GGAGTGGGAGGCAAACC 18 502 740321 N/A N/A3446 3462 CGGCGACGCGAGGCTGG 16 503 740322 N/A N/A 3449 3465AGCCGGCGACGCGAGGC 25 504 740323 N/A N/A 3452 3468 GTGAGCCGGCGACGCGA 51505 740324 N/A N/A 3455 3471 GCTGTGAGCCGGCGACG 66 506 740325 N/A N/A3458 3474 GCCGCTGTGAGCCGGCG  5 507 740326 N/A N/A 3463 3479AGGAGGCCGCTGTGAGC 45 508 740327 N/A N/A 3466 3482 CAGAGGAGGCCGCTGTG 21509 740328 N/A N/A 3469 3485 CCCCAGAGGAGGCCGCT 23 510 740329 N/A N/A3472 3488 TGTCCCCAGAGGAGGCC  0 511 740330 N/A N/A 3475 3491GACTGTCCCCAGAGGAG 24 512 740331 N/A N/A 3496 3512 GCGGAGGCGGCACCCGG 13513 740332 N/A N/A 3499 3515 AGGGCGGAGGCGGCACC 25 514 740333 N/A N/A3502 3518 GGAAGGGCGGAGGCGGC 28 515 740334 N/A N/A 3505 3521ACAGGAAGGGCGGAGGC  1 516 740335 N/A N/A 3508 3524 CGCACAGGAAGGGCGGA 19517 740336 N/A N/A 3511 3527 GAGCGCACAGGAAGGGC 33 518 740337 N/A N/A3514 3530 AAGGAGCGCACAGGAAG 64 519 740338 N/A N/A 3518 3534GGAAAAGGAGCGCACAG 40 520 740339 N/A N/A 3521 3537 GAAGGAAAAGGAGCGCA 36521 740340 N/A N/A 3532 3548 ATAGGAAAGAAGAAGGA 42 522 740341 N/A N/A3536 3552 TTTAATAGGAAAGAAGA  3 523 740342 N/A N/A 3540 3556AATATTTAATAGGAAAG  0 524 740343 N/A N/A 3548 3564 TCCCAAATAATATTTAA 42525 740344 N/A N/A 3551 3567 AATTCCCAAATAATATT 28 526 740345 N/A N/A3554 3570 AACAATTCCCAAATAAT 33 527 740346 N/A N/A 3558 3574TTTAAACAATTCCCAAA 15 528 740347 N/A N/A 3561 3577 AAATTTAAACAATTCCC 48529 740348 N/A N/A 3587 3603 CCCGCCTCTCTCTTTTT 20 530 740349 N/A N/A3590 3606 CTCCCCGCCTCTCTCTT  0 531 740350 N/A N/A 3594 3610ACTCCTCCCCGCCTCTC  2 532 740351 N/A N/A 3598 3614 TCCGACTCCTCCCCGCC 40533 740352 N/A N/A 3601 3617 AACTCCGACTCCTCCCC 55 534 740353 N/A N/A3604 3620 CACAACTCCGACTCCTC 56 535 740354 N/A N/A 3607 3623CTCCACAACTCCGACTC 31 536 740355 N/A N/A 3610 3626 CTTCTCCACAACTCCGA 41537 740356 N/A N/A 3613 3629 CTGCTTCTCCACAACTC 27 538 740357 N/A N/A3616 3632 CCTCTGCTTCTCCACAA 30 539 740358 N/A N/A 3620 3636AGTCCCTCTGCTTCTCC  0 540 740359 N/A N/A 3623 3639 CTGAGTCCCTCTGCTTC 26541 740369  10  26 3192 3208 CTAGTCCTCCTCCTTCT 14 542 740370  23  393205 3221 CGTCCTCCTCCTCCTAG 27 543 740371  28  44 3210 3226GTCGCCGTCCTCCTCCT  0 544 740372  45  61 3227 3243 TTGGGCCCCTTCTGGTC  0545 740373  48  64 3230 3246 CTCTTGGGCCCCTTCTG 42 546 740374  51  673233 3249 CCTCTCTTGGGCCCCTT 43 547 740375  54  70 3236 3252CCCCCTCTCTTGGGCCC 31 548 740376  57  73 3239 3255 TCGCCCCCTCTCTTGGG  0549 740377  60  76 3242 3258 CGCTCGCCCCCTCTCTT 23 550 740378  63  793245 3261 GGTCGCTCGCCCCCTCT 35 551 740379  67  83 3249 3265GCTCGGTCGCTCGCCCC 53 552 740380  92 108 3274 3290 ACGCACCTCACTTCCGC 58553 740381  95 111 3277 3293 CGCACGCACCTCACTTC 43 554 740382  98 1143280 3296 GCCCGCACGCACCTCAC 42 555 740383 101 117 3283 3299GCAGCCCGCACGCACCT 44 556 740384 104 120 3286 3302 GCTGCAGCCCGCACGCA 19557 740385 107 123 3289 3305 TGCGCTGCAGCCCGCAC  4 558 740386 110 1263292 3308 GTCTGCGCTGCAGCCCG 59 559 740387 169 185 3351 3367AGGCTTGAAGGCAAGGC 69 560 740388 172 188 3354 3370 AGAAGGCTTGAAGGCAA 66561 740389 175 191 3357 3373 GGCAGAAGGCTTGAAGG 44 562 740390 178 1943360 3376 AAAGGCAGAAGGCTTGA 44 563 740391 181 197 3363 3379TGGAAAGGCAGAAGGCT 59 564 740392 184 200 3366 3382 GGGTGGAAAGGCAGAAG 32565

TABLE 9Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 387978 282 301 4733 4752TCCTTGGCCTTTGAAAGTCC  9  21 740410 240 256 4691 4707 GAATTCCTTTACACCAC86  33 740393 187 203 3369 3385 CGAGGGTGGAAAGGCAG 56 566 740394 191 2073373 3389 CTCACGAGGGTGGAAAG 30 567 740395 194 210 3376 3392CCGCTCACGAGGGTGGA 57 568 740396 197 213 3379 3395 TCTCCGCTCACGAGGGT 40569 740397 200 216 3382 3398 AGTTCTCCGCTCACGAG 52 570 740398 203 2193385 3401 CCCAGTTCTCCGCTCAC 43 571 740399 206 222 3388 3404ACTCCCAGTTCTCCGCT 33 572 740400 209 225 3391 3407 GCCACTCCCAGTTCTCC 44573 740401 213 229 3395 3411 AATGGCCACTCCCAGTT 38 574 740402 216 2323398 3414 TCGAATGGCCACTCCCA 45 575 740403 233 249 N/A N/ATTTACACCACACTGTCG 50 108 740404 234 250 N/A N/A CTTTACACCACACTGTC 62 577740405 235 251 N/A N/A CCTTTACACCACACTGT 77 576 740406 236 252 4687 4703TCCTTTACACCACACTG 85 260 740407 237 253 4688 4704 TTCCTTTACACCACACT 88335 740408* 238 254 4689 4705 ATTCCTTTACACCACAC 83 412 740409* 239 2554690 4706 AATTCCTTTACACCACA 89 577 740411 241 257 4692 4708TGAATTCCTTTACACCA 83 578 740412 242 258 4693 4709 ATGAATTCCTTTACACC 87584 740413 243 259 4694 4710 AATGAATTCCTTTACAC 78 579 740414 245 2614696 4712 CTAATGAATTCCTTTAC 82 580 740415 246 262 4697 4713GCTAATGAATTCCTTTA 80 581 740416 249 265 4700 4716 ATGGCTAATGAATTCCT 89582 740417 253 269 4704 4720 ATCCATGGCTAATGAAT 64 583 740418 254 2704705 4721 CATCCATGGCTAATGAA 69 336 740419 255 271 4706 4722ACATCCATGGCTAATGA 76 413 740420 256 272 4707 4723 TACATCCATGGCTAATG 73 34 740421 257 273 4708 4724 ATACATCCATGGCTAAT 74 593 740422 258 2744709 4725 AATACATCCATGGCTAA 85 186 740423 259 275 4710 4726GAATACATCCATGGCTA 76 584 740424 260 276 4711 4727 TGAATACATCCATGGCT 77262 740425 261 277 4712 4728 ATGAATACATCCATGGC 83 585 740426 263 2794714 4730 TCATGAATACATCCATG 55 586 740427 265 281 4716 4732TTTCATGAATACATCCA 88 587 740428 266 282 4717 4733 CTTTCATGAATACATCC 76414 740429 267 283 4718 4734 CCTTTCATGAATACATC 86 588 740430 268 2844719 4735 TCCTTTCATGAATACAT 91 589 740431 269 285 4720 4736GTCCTTTCATGAATACA 82 590 740432 270 286 4721 4737 AGTCCTTTCATGAATAC 92591 740433 271 287 4722 4738 AAGTCCTTTCATGAATA 68 592 740434 273 2894724 4740 GAAAGTCCTTTCATGAA 67 593 740435 275 291 4726 4742TTGAAAGTCCTTTCATG 25 594 740436 276 292 4727 4743 TTTGAAAGTCCTTTCAT  8595 740437 277 293 4728 4744 CTTTGAAAGTCCTTTCA 66 596 740438 278 2944729 4745 CCTTTGAAAGTCCTTTC 86  35 740439 279 295 4730 4746GCCTTTGAAAGTCCTTT 88 597 740440 280 296 4731 4747 GGCCTTTGAAAGTCCTT 88111 740441 281 297 4732 4748 TGGCCTTTGAAAGTCCT 58 598 740442 282 2984733 4749 TTGGCCTTTGAAAGTCC 68 187 740443 283 299 4734 4750CTTGGCCTTTGAAAGTC 75 263 740444 285 301 4736 4752 TCCTTGGCCTTTGAAAG 47599 740445 286 302 4737 4753 CTCCTTGGCCTTTGAAA 57 415 740446 301 3174752 4768 AGCAGCCACAACTCCCT 62 112 740447 302 318 4753 4769CAGCAGCCACAACTCCC 65 600 740448 304 320 4755 4771 AGCAGCAGCCACAACTC 63601 740449 305 321 4756 4772 CAGCAGCAGCCACAACT 52 602 740450 308 3244759 4775 TCTCAGCAGCAGCCACA 63 603 740451 309 325 4760 4776TTCTCAGCAGCAGCCAC 69 604 740452 311 327 4762 4778 TTTTCTCAGCAGCAGCC 75605 740453 312 328 4763 4779 GTTTTCTCAGCAGCAGC 66 188 740454 313 3294764 4780 GGTTTTCTCAGCAGCAG 79 606 740455 314 330 4765 4781TGGTTTTCTCAGCAGCA 78 607 740456 317 333 4768 4784 GTTTGGTTTTCTCAGCA 82608 740457 326 342 4777 4793 CCACACCCTGTTTGGTT 71 609 740458 329 3454780 4796 CTGCCACACCCTGTTTG 54 610 740459 332 348 4783 4799CTTCTGCCACACCCTGT 74 611 740460 333 349 4784 4800 GCTTCTGCCACACCCTG 73612 740461 335 351 4786 4802 CTGCTTCTGCCACACCC 80 613 740462 336 3524787 4803 GCTGCTTCTGCCACACC 77 339 740463 338 354 4789 4805CTGCTGCTTCTGCCACA 64 614 740464 339 355 4790 4806 CCTGCTGCTTCTGCCAC 52615 740465 342 358 4793 4809 TTTCCTGCTGCTTCTGC 63 616 740466 345 3614796 4812 GTCTTTCCTGCTGCTTC 69 617 740467 348 364 4799 4815TTTGTCTTTCCTGCTGC 56 618 740468 362 378 4813 4829 AGAGAACACCCTCTTTT 47619 740469 365 381 4816 4832 CATAGAGAACACCCTCT 71 620 740470 368 3844819 4835 CTACATAGAGAACACCC 81 621

TABLE 10Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 387978 282 301  4733  4752TCCTTGGCCTTTGAAAGTCC 32  21 740410 240 256  4691  4707 GAATTCCTTTACACCAC90  33 740471 369 385  4820  4836 CCTACATAGAGAACACC 76 622 740472 371387 N/A N/A AGCCTACATAGAGAACA 51 623 740473 372 388 N/A N/AGAGCCTACATAGAGAAC 74  37 740474 373 389 N/A N/A GGAGCCTACATAGAGAA 65 624740475 374 390 N/A N/A TGGAGCCTACATAGAGA 46 625 740476 375 391 N/A N/ATTGGAGCCTACATAGAG 61 626 740477 378 394 N/A N/A GTTTTGGAGCCTACATA 56 627740478 379 395 N/A N/A GGTTTTGGAGCCTACAT 72 189 740479 380 396 1219312209 TGGTTTTGGAGCCTACA 78 628 740480 381 397 12194 12210TTGGTTTTGGAGCCTAC 64 265 740481 382 398 12195 12211 CTTGGTTTTGGAGCCTA 43340 740482 383 399 12196 12212 CCTTGGTTTTGGAGCCT 81 417 740483 384 40012197 12213 TCCTTGGTTTTGGAGCC 79 629 740484 385 401 12198 12214CTCCTTGGTTTTGGAGC 21  38 740485 386 402 12199 12215 CCTCCTTGGTTTTGGAG 19630 740486 388 404 12201 12217 TCCCTCCTTGGTTTTGG 63 631 740487 391 40712204 12220 CACTCCCTCCTTGGTTT 71 632 740488 396 412 12209 12225TGCACCACTCCCTCCTT 62 633 740489 399 415 12212 12228 CCATGCACCACTCCCTC 51634 740490 400 416 12213 12229 ACCATGCACCACTCCCT 61 266 740491 401 41712214 12230 CACCATGCACCACTCCC 80 635 740492 402 418 12215 12231ACACCATGCACCACTCC 69 341 740493 403 419 12216 12232 CACACCATGCACCACTC 69636 740494 404 420 12217 12233 CCACACCATGCACCACT 78 418 740495 406 42212219 12235 TGCCACACCATGCACCA 75 646 740496 407 423 12220 12236TTGCCACACCATGCACC 68 637 740497 408 424 12221 12237 GTTGCCACACCATGCAC 50191 740498 409 425 12222 12238 TGTTGCCACACCATGCA 81 638 740499 410 42612223 12239 CTGTTGCCACACCATGC 79 267 740500 411 427 N/A N/AACTGTTGCCACACCATG 88 639 740501 418 434 N/A N/A CTCAGCCACTGTTGCCA 68 342740502 419 435 N/A N/A TCTCAGCCACTGTTGCC 66 640 740503 421 437 N/A N/ACTTCTCAGCCACTGTTG 57 641 740504 422 438 N/A N/A TCTTCTCAGCCACTGTT 59 642740505 427 443 17994 18010 TTTGGTCTTCTCAGCCA 41 643 740506 432 448 1799918015 TGCTCTTTGGTCTTCTC 27 644 740507 435 451 18002 18018ACTTGCTCTTTGGTCTT 66 645 740508 437 453 18004 18020 TCACTTGCTCTTTGGTC 83646 740509 438 454 18005 18021 GTCACTTGCTCTTTGGT 89 647 740510 439 45518006 18022 TGTCACTTGCTCTTTGG 87 648 740511 440 456 18007 18023TTGTCACTTGCTCTTTG 79  40 740512 441 457 18008 18024 TTTGTCACTTGCTCTTT 72649 740513 442 458 18009 18025 ATTTGTCACTTGCTCTT 82 116 740514 443 45918010 18026 CATTTGTCACTTGCTCT 76 650 740515 444 460 18011 18027ACATTTGTCACTTGCTC 80 192 740516 445 461 18012 18028 AACATTTGTCACTTGCT 80268 740517 446 462 18013 18029 CAACATTTGTCACTTGC 86 343 740518 447 46318014 18030 CCAACATTTGTCACTTG 48 651 740519 448 464 18015 18031TCCAACATTTGTCACTT 40 420 740520 449 465 18016 18032 CTCCAACATTTGTCACT 56652 740521 451 467 18018 18034 TCCTCCAACATTTGTCA 17 653 740522 454 47018021 18037 TGCTCCTCCAACATTTG 49 654 740523 457 473 18024 18040CACTGCTCCTCCAACAT 60 655 740524 460 476 18027 18043 CACCACTGCTCCTCCAA 81656 740525 463 479 18030 18046 CGTCACCACTGCTCCTC 55 657 740526 464 48018031 18047 CCGTCACCACTGCTCCT 69 193 740527 466 482 18033 18049ACCCGTCACCACTGCTC 87 658 740528 467 483 18034 18050 CACCCGTCACCACTGCT 82344 740529 468 484 18035 18051 ACACCCGTCACCACTGC 76 659 740530 470 48618037 18053 TCACACCCGTCACCACT 77 681 740531 471 487 18038 18054GTCACACCCGTCACCAC 79 118 740532 472 488 18039 18055 TGTCACACCCGTCACCA 72660 740533 473 489 18040 18056 CTGTCACACCCGTCACC 88 194 740534 474 49018041 18057 GCTGTCACACCCGTCAC 84 661 740535 476 492 18043 18059CTGCTGTCACACCCGTC 85 662 740536 479 495 18046 18062 CTACTGCTGTCACACCC 75663 740537 482 498 18049 18065 GGGCTACTGCTGTCACA 59 664 740538 485 50118052 18068 TCTGGGCTACTGCTGTC 54 665 740539 488 504 18055 18071TCTTCTGGGCTACTGCT 48 666 740540 491 507 18058 18074 CTGTCTTCTGGGCTACT 61667 740541 494 510 18061 18077 CCACTGTCTTCTGGGCT 61 668 740542 498 51418065 18081 CCCTCCACTGTCTTCTG 26 669 740543 502 518 18069 18085TGCTCCCTCCACTGTCT 62 670 740544 510 526 18077 18093 ATGCTCCCTGCTCCCTC 70671 740545 513 529 18080 18096 GCAATGCTCCCTGCTCC 88 672 740546 523 53918090 18106 AGTGGCTGCTGCAATGC 61 119 740547 526 542 18093 18109GCCAGTGGCTGCTGCAA 58 673

TABLE 11Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 387978 282 301   4733   4752TCCTTGGCCTTTGAAAGTCC 11  21 740410 240 256   4691   4707GAATTCCTTTACACCAC 89  33 740548 529 545  18096  18112 AAAGCCAGTGGCTGCTG76 674 740549 532 548  18099  18115 GACAAAGCCAGTGGCTG 72 675 740550 535551  18102  18118 TTTGACAAAGCCAGTGG 63 676 740551 538 554  18105  18121CTTTTTGACAAAGCCAG 71 677 740552 541 557  18108  18124 GTCCTTTTTGACAAAGC31 678 740553 544 560  18111  18127 CTGGTCCTTTTTGACAA 50 679 740554 547563  18114  18130 CAACTGGTCCTTTTTGA 67 680 740555 550 566  18117  18133GCCCAACTGGTCCTTTT 73 681 740556 553 569  18120  18136 CTTGCCCAACTGGTCCT55 682 740557 557 573 N/A N/A CATTCTTGCCCAACTGG 15 683 740558 560 576N/A N/A CTTCATTCTTGCCCAAC 60 684 740559 563 579 N/A N/ACTTCTTCATTCTTGCCC 72 685 740560 566 582 N/A N/A CTCCTTCTTCATTCTTG 48 686740561 569 585 111104 111120 GGGCTCCTTCTTCATTC 60 687 740562 585 601111120 111136 AGAATTCCTTCCTGTGG 38 688 740563 588 604 111123 111139TCCAGAATTCCTTCCTG 63 689 740564 591 607 111126 111142 TCTTCCAGAATTCCTTC45 690 740565 594 610 111129 111145 ATATCTTCCAGAATTCC 63 691 740566 597613 111132 111148 GGCATATCTTCCAGAAT 73 692 740567 600 616 111135 111151ACAGGCATATCTTCCAG 48 693 740568 603 619 111138 111154 TCCACAGGCATATCTTC46 694 740569 607 623 111142 111158 AGGATCCACAGGCATAT 34 695 740570 610626 111145 111161 GTCAGGATCCACAGGCA 72 696 740571 613 629 111148 111164ATTGTCAGGATCCACAG 21 697 740572 616 632 111151 111167 CTCATTGTCAGGATCCA75 698 740573 619 635 111154 111170 AGCCTCATTGTCAGGAT 79 699 740574 622638 111157 111173 ATAAGCCTCATTGTCAG 31 700 740575 625 641 111160 111176TTCATAAGCCTCATTGT  0 701 740576 627 643 111162 111178 ATTTCATAAGCCTCATT35 702 740577 629 645 111164 111180 GCATTTCATAAGCCTCA 78 703 740578 632648 111167 111183 AAGGCATTTCATAAGCC 67 704 740579 635 651 111170 111186CAGAAGGCATTTCATAA 70 705 740580 638 654 111173 111189 CCTCAGAAGGCATTTCA31 706 740581 641 657 N/A N/A CTTCCTCAGAAGGCATT 62 707 740582 644 660N/A N/A ACCCTTCCTCAGAAGGC 60 708 740583 647 663 N/A N/AGATACCCTTCCTCAGAA  4 709 740584 651 667 N/A N/A TCTTGATACCCTTCCTC 29 710740585 654 670 113722 113738 TAGTCTTGATACCCTTC 70 711 740586 672 688113740 113756 TCTTAGGCTTCAGGTTC 66 712 740587 675 691 113743 113759ATTTCTTAGGCTTCAGG 47 713 740588 678 694 113746 113762 GATATTTCTTAGGCTTC61 714 740589 681 697 113749 113765 AAAGATATTTCTTAGGC 43 715 740590 684700 113752 113768 AGCAAAGATATTTCTTA 49 716 740591 687 703 113755 113771GGGAGCAAAGATATTTC 80 717 740592 690 706 113758 113774 ACTGGGAGCAAAGATAT55 718 740593 694 710 113762 113778 AGAAACTGGGAGCAAAG 86 719 740594 697713 113765 113781 TCAAGAAACTGGGAGCA 49 720 740595 700 716 113768 113784ATCTCAAGAAACTGGGA 69 721 740596 703 719 113771 113787 CAGATCTCAAGAAACTG72 722 740597 706 722 113774 113790 CAGCAGATCTCAAGAAA 72 723 740598 709725 113777 113793 TGTCAGCAGATCTCAAG 47 724 740599 712 728 113780 113796ATCTGTCAGCAGATCTC 32 725 740600 716 732 113784 113800 GAACATCTGTCAGCAGA 0 726 740601 719 735 113787 113803 ATGGAACATCTGTCAGC  9 727 740602 722738 113790 113806 AGGATGGAACATCTGTC 19 728 740603 725 741 113793 113809TACAGGATGGAACATCT  0 729 740604 730 746 113798 113814 ACTTGTACAGGATGGAA55 730 740605 733 749 113801 113817 AGCACTTGTACAGGATG 61 731 740606 736752 113804 113820 CTGAGCACTTGTACAGG 66 732 740607 739 755 113807 113823GAACTGAGCACTTGTAC 49 733 740608 742 758 113810 113826 TTGGAACTGAGCACTTG41 734 740609 745 761 113813 113829 ACATTGGAACTGAGCAC 36 735 740610 748764 113816 113832 GGCACATTGGAACTGAG 47 736 740611 752 768 113820 113836ACTGGGCACATTGGAAC 51 737 740612 755 771 113823 113839 ATGACTGGGCACATTGG44 738 740613 758 774 113826 113842 GTCATGACTGGGCACAT 38 739 740614 761777 113829 113845 AATGTCATGACTGGGCA 32 740 740615 764 780 113832 113848AGAAATGTCATGACTGG 76 741 740616 767 783 113835 113851 TTGAGAAATGTCATGAC54 742 740617 770 786 113838 113854 ACTTTGAGAAATGTCAT 34 743 740618 773789 113841 113857 AAAACTTTGAGAAATGT 30 744 740619 776 792 113844 113860GTAAAAACTTTGAGAAA 69 745 740620 779 795 113847 113863 ACTGTAAAAACTTTGAG64 746 740621 782 798 113850 113866 TACACTGTAAAAACTTT 39 747 740622 785801 113853 113869 AGATACACTGTAAAAAC 27 748 740623 786 802 113854 113870GAGATACACTGTAAAAA 36 749 740624 787 803 113855 113871 CGAGATACACTGTAAAA56 750

TABLE 12Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 387978  282  301   4733   4752TCCTTGGCCTTTGAAAGTCC 11  21 740410  240  256   4691   4707GAATTCCTTTACACCAC 89  33 740625  791  807 113859 113875ACTTCGAGATACACTGT 76 751 740626  793  809 113861 113877AGACTTCGAGATACACT 72 275 740627  795  811 113863 113879GAAGACTTCGAGATACA 63 752 740628  796  812 113864 113880GGAAGACTTCGAGATAC 71 427 740629  797  813 113865 113881TGGAAGACTTCGAGATA 31 753 740630  799  815 113867 113883GATGGAAGACTTCGAGA 50 754 740631  802  818 113870 113886GCTGATGGAAGACTTCG 67 755 740632  805  821 113873 113889ACTGCTGATGGAAGACT 73 756 740633  808  824 113876 113892ATCACTGCTGATGGAAG 55 757 740634  812  828 113880 113896TTCAATCACTGCTGATG 15 758 740635  815  831 113883 113899TACTTCAATCACTGCTG 60 759 740636  818  834 113886 113902AGATACTTCAATCACTG 72 760 740637  819  835 113887 113903CAGATACTTCAATCACT 48 761 740638  820  836 113888 113904ACAGATACTTCAATCAC 60 762 740639  821  837 113889 113905TACAGATACTTCAATCA 38 763 740640  824  840 113892 113908AGGTACAGATACTTCAA 63 764 740641  827  843 113895 113911GGCAGGTACAGATACTT 45 765 740642  845  861 113913 113929ACCGAAATGCTGAGTGG 63 766 740643  848  864 113916 113932AGCACCGAAATGCTGAG 73 767 740644  851  867 113919 113935GGAAGCACCGAAATGCT 48 768 740645  854  870 113922 113938AAGGGAAGCACCGAAAT 46 769 740646  857  873 113925 113941TGAAAGGGAAGCACCGA 34 770 740647  860  876 113928 113944CAGTGAAAGGGAAGCAC 72 771 740648  863  879 113931 113947CTTCAGTGAAAGGGAAG 21 772 740649  865  881 113933 113949CACTTCAGTGAAAGGGA 75 773 740650  866  882 113934 113950TCACTTCAGTGAAAGGG 79  49 740651  867  883 113935 113951TTCACTTCAGTGAAAGG 31 774 740652  869  885 113937 113953TATTCACTTCAGTGAAA  0 775 740653  870  886 113938 113954GTATTCACTTCAGTGAA 35 776 740654  873  889 113941 113957CATGTATTCACTTCAGT 78 777 740655  876  892 113944 113960TACCATGTATTCACTTC 67 778 740656  879  895 113947 113963TGCTACCATGTATTCAC 70 779 740657  882  898 113950 113966CCCTGCTACCATGTATT 31 780 740658  885  901 113953 113969AGACCCTGCTACCATGT 62 781 740659  886  902 113954 113970AAGACCCTGCTACCATG 60 782 740660  890  906 113958 113974CACAAAGACCCTGCTAC  4 783 740661  892  908 113960 113976CACACAAAGACCCTGCT 29  50 740662  894  910 113962 113978AGCACACAAAGACCCTG 70 784 740663  895  911 113963 113979CAGCACACAAAGACCCT 66 202 740664  896  912 113964 113980ACAGCACACAAAGACCC 47 785 740665  898  914 113966 113982CCACAGCACACAAAGAC 61 786 740666  901  917 113969 113985AATCCACAGCACACAAA 43 787 740667  905  921 113973 113989ACAAAATCCACAGCACA 49 788 740668  911  927 113979 113995GAAGCCACAAAATCCAC 80 789 740669  915  931 113983 113999GATTGAAGCCACAAAAT 55 790 740670  918  934 113986 114002GTAGATTGAAGCCACAA 86 791 740671  935  951 114003 114019TAATTTGTTTTAACATC 49 792 740672  943  959 114011 114027GGTGTTTTTAATTTGTT 69 793 740673  944  960 114012 114028AGGTGTTTTTAATTTGT 72 128 740674  945  961 114013 114029TAGGTGTTTTTAATTTG 72 204 740675  946  962 114014 114030TTAGGTGTTTTTAATTT 47 794 740676  947  963 114015 114031CTTAGGTGTTTTTAATT 32 280 740677  950  966 114018 114034TCACTTAGGTGTTTTTA  0 795 740678  953  969 114021 114037TAGTCACTTAGGTGTTT  9 796 740679  957  973 114025 114041GTGGTAGTCACTTAGGT 19 797 740680  960  976 114028 114044TAAGTGGTAGTCACTTA  0 798 740681  963  979 114031 114047AAATAAGTGGTAGTCAC 55 799 740682  966  982 114034 114050TAGAAATAAGTGGTAGT 61 800 740683  969  985 114037 114053ATTTAGAAATAAGTGGT 66 801 740684  972  988 114040 114056AGGATTTAGAAATAAGT 49 802 740685  975  991 114043 114059GTGAGGATTTAGAAATA 41 803 740686  976  992 114044 114060AGTGAGGATTTAGAAAT 36 804 740687  977  993 114045 114061TAGTGAGGATTTAGAAA 47 805 740688  979  995 114047 114063AATAGTGAGGATTTAGA 51 806 740689  982  998 114050 114066AAAAATAGTGAGGATTT 44 807 740690  985 1001 114053 114069CAAAAAAATAGTGAGGA 38 808 740691  989 1005 114057 114073GCAACAAAAAAATAGTG 32 809 740692 1002 1018 114070 114086CTTCTGAACAACAGCAA 76 810 740693 1005 1021 114073 114089CAACTTCTGAACAACAG 54 811 740694 1008 1024 114076 114092TAACAACTTCTGAACAA 34 812 740695 1011 1027 114079 114095CACTAACAACTTCTGAA 30 813 740696 1014 1030 114082 114098AATCACTAACAACTTCT 69 814 740697 1017 1033 114085 114101GCAAATCACTAACAACT 64 815 740698 1020 1036 114088 114104ATAGCAAATCACTAACA 39 816 740699 1024 1040 114092 114108TATGATAGCAAATCACT 27 817 740700 1027 1043 114095 114111ATATATGATAGCAAATC 36 818 740701 1030 1046 114098 114114ATAATATATGATAGCAA 56 819 740470  368  384   4819   4835CTACATAGAGAACACCC 81 621

TABLE 13Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 387978  282  301   4733   4752TCCTTGGCCTTTGAAAGTCC 18  21 740410  240  256   4691   4707GAATTCCTTTACACCAC 84  33 740702 1033 1049 114101 114117CTTATAATATATGATAG 16 820 740703 1036 1052 114104 114120AATCTTATAATATATGA 27 821 740704 1041 1057 114109 114125CTAAAAATCTTATAATA  0 822 740705 1044 1060 114112 114128CACCTAAAAATCTTATA 11 823 740706 1047 1063 114115 114131AGACACCTAAAAATCTT 49 824 740707 1048 1064 114116 114132AAGACACCTAAAAATCT 25 825 740708 1049 1065 114117 114133AAAGACACCTAAAAATC 27 826 740709 1052 1068 114120 114136TTAAAAGACACCTAAAA  6 827 740710 1055 1071 114123 114139TCATTAAAAGACACCTA 30 828 740711 1058 1074 114126 114142GTATCATTAAAAGACAC 30 829 740712 1061 1077 114129 114145ACAGTATCATTAAAAGA 35 830 740713 1064 1080 114132 114148TAGACAGTATCATTAAA 54 831 740714 1068 1084 114136 114152TTCTTAGACAGTATCAT 57 832 740715 1071 1087 114139 114155TTATTCTTAGACAGTAT 64 833 740716 1074 1090 114142 114158TCATTATTCTTAGACAG 47 834 740717 1092 1108 114160 114176AACAAATTTCACAATAC 60 835 740718 1095 1111 114163 114179ATTAACAAATTTCACAA 34 836 740719 1106 1122 114174 114190GTATTATATATATTAAC 32 837 740720 1121 1137 114189 114205CTCACATATTTTTAAGT 44 838 740721 1124 1140 114192 114208ATGCTCACATATTTTTA 45 839 740722 1127 1143 114195 114211TTCATGCTCACATATTT 40 840 740723 1130 1146 114198 114214AGTTTCATGCTCACATA 67 841 740724 1134 1150 114202 114218GCATAGTTTCATGCTCA 51 842 740725 1137 1153 114205 114221GGTGCATAGTTTCATGC 46 843 740726 1138 1154 114206 114222AGGTGCATAGTTTCATG 54 844 740727 1140 1156 114208 114224ATAGGTGCATAGTTTCA 63 845 740728 1143 1159 114211 114227TTTATAGGTGCATAGTT 62 846 740729 1146 1162 114214 114230GTATTTATAGGTGCATA 67 847 740730 1149 1165 114217 114233TTAGTATTTATAGGTGC 72 848 740731 1152 1168 114220 114236TATTTAGTATTTATAGG 32 849 740732 1164 1180 114232 114248GGTAAAATTTCATATTT 33 850 740733 1173 1189 114241 114257TCGCAAAATGGTAAAAT 49 851 740734 1176 1192 114244 114260ACATCGCAAAATGGTAA 62 852 740735 1179 1195 114247 114263AACACATCGCAAAATGG 42 853 740736 1182 1198 114250 114266TAAAACACATCGCAAAA 28 854 740737 1185 1201 114253 114269GAATAAAACACATCGCA 62 855 740738 1188 1204 114256 114272AGTGAATAAAACACATC 16 856 740739 1191 1207 114259 114275ACAAGTGAATAAAACAC 64 857 740740 1196 1212 114264 114280CAAACACAAGTGAATAA 16 858 740741 1199 1215 114267 114283ATACAAACACAAGTGAA 33 859 740742 1203 1219 114271 114287TTATATACAAACACAAG 31 860 740743 1207 1223 114275 114291CCATTTATATACAAACA 28 861 740744 1210 1226 114278 114294TCACCATTTATATACAA 53 862 740745 1214 1230 114282 114298ATTCTCACCATTTATAT 40 863 740746 1222 1238 114290 114306TTATTTTAATTCTCACC 46 864 740747 1242 1258 114310 114326TTTTGCAATGAGATAAC  0 865 740748 1245 1261 114313 114329TATTTTTGCAATGAGAT 42 866 740749 1265 1281 114333 114349GAGATGGGATAAAAATA 38 867 740750 1268 1284 114336 114352AGTGAGATGGGATAAAA 32 868 740751 1271 1287 114339 114355TAAAGTGAGATGGGATA 30 869 740752 1275 1291 114343 114359TTATTAAAGTGAGATGG 24 870 740753 1278 1294 114346 114362TTATTATTAAAGTGAGA  9 871 740754 1288 1304 114356 114372AGCATGATTTTTATTAT 36 872 740755 1291 1307 114359 114375ATAAGCATGATTTTTAT  2 873 740756 1292 1308 114360 114376TATAAGCATGATTTTTA 25 874 740757 1296 1312 114364 114380TGCTTATAAGCATGATT 20 875 740758 1299 1315 114367 114383TGTTGCTTATAAGCATG  0 876 740759 1302 1318 114370 114386TCATGTTGCTTATAAGC 27 877 740760 1306 1322 114374 114390TAATTCATGTTGCTTAT 55 878 740761 1309 1325 114377 114393TCTTAATTCATGTTGCT 35 879 740762 1312 1328 114380 114396AGTTCTTAATTCATGTT 41 880 740763 1315 1331 114383 114399GTCAGTTCTTAATTCAT 54 881 740764 1318 1334 114386 114402TGTGTCAGTTCTTAATT 61 882 740765 1321 1337 114389 114405CTTTGTGTCAGTTCTTA 68 883 740766 1324 1340 114392 114408GTCCTTTGTGTCAGTTC 64 884 740767 1328 1344 114396 114412TTTTGTCCTTTGTGTCA 30 885 740768 1331 1347 114399 114415TATTTTTGTCCTTTGTG 36 886 740769 1336 1352 114404 114420CTTTATATTTTTGTCCT 13 887 740770 1346 1362 114414 114430CTATTAATAACTTTATA 15 888 740771 1349 1365 114417 114433TGGCTATTAATAACTTT 43 889 740772 1352 1368 114420 114436AAATGGCTATTAATAAC 36 890 740773 1355 1371 114423 114439TTCAAATGGCTATTAAT 35 891 740774 1358 1374 114426 114442TTCTTCAAATGGCTATT 40 892 740775 1361 1377 114429 114445TCCTTCTTCAAATGGCT 45 893 740776 1364 1380 114432 114448TCCTCCTTCTTCAAATG  8 894 740777 1369 1385 114437 114453AAAATTCCTCCTTCTTC 39 895 740778 1372 1388 114440 114456TCTAAAATTCCTCCTTC 33 896

Example 3: Effect of 4-9-4 MOE and cEt Gapmers with MixedInternucleoside Linkages on Human SNCA In Vitro, Single Dose

Modified oligonucleotides complementary to a human SNCA nucleic acidwere designed and tested as described in Example 1 for their effect onSNCA mRNA in vitro. The modified oligonucleotides were tested in aseries of experiments that had similar culture conditions.

The modified oligonucleotides in tables 14-23 are 4-9-4 MOE and cEtgapmers. The gapmers are 17 nucleobases in length, wherein the centralgap segment comprises nine 2′-deoxynucleosides and is flanked by wingsegments on both the 5′ end on the 3′ end comprising two 2′-MOEnucleosides and two cEt nucleosides. The sugar motif for the gapmers is(from 5′ to 3′): eekkdddddddddkkee; wherein ‘d’ represents a2′-deoxyribose sugar; ‘e’ represents a 2′-MOE modified sugar; and ‘k’represents a cEt modified sugar. All cytosine residues throughout eachgapmer are 5-methyl cytosines. The internucleoside linkages are mixedphosphodiester and phosphorothioate linkages. The internucleosidelinkage motif for the gapmers is (from 5′ to 3′): sooosssssssssoss;wherein ‘o’ represents a phosphodiester internucleoside linkage and ‘s’represents a phosphorothioate internucleoside linkage. “Start Site”indicates the 5′-most nucleoside to which the gapmer is complementary inthe human nucleic acid sequence. “Stop Site” indicates the 3′-mostnucleoside to which the gapmer is complementary in the human nucleicacid sequence.

Each modified oligonucleotide listed in the Tables below iscomplementary to human SNCA nucleic acid sequences SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6, asindicated. ‘N/A’ indicates that the modified oligonucleotide is notcomplementary to that particular nucleic acid with 100% complementarity.A value of 0% reduction indicates that the compound had no effect orincreased mRNA concentrations in the cell. As shown below, modifiedoligonucleotides complementary to human SNCA reduced the amount of humanSNCA mRNA.

TABLE 14Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 740410  240  256   4691   4707GAATTCCTTTACACCAC 97  33 740432  270  286   4721   4737AGTCCTTTCATGAATAC 95 591 740779 1375 1391 114443 114459TCTTCTAAAATTCCTCC 51 897 740780 1378 1394 114446 114462ACCTCTTCTAAAATTCC 45 898 740781 1381 1397 114449 114465TCTACCTCTTCTAAAAT  7 899 740782 1384 1400 114452 114468TTCTCTACCTCTTCTAA 41 900 740783 1389 1405 114457 114473CCATTTTCTCTACCTCT 90 901 740784 1392 1408 114460 114476GTTCCATTTTCTCTACC 78 902 740785 1396 1412 114464 114480TAATGTTCCATTTTCTC 62 903 740786 1400 1416 114468 114484GGGTTAATGTTCCATTT 65 904 740787 1403 1419 114471 114487GTAGGGTTAATGTTCCA 74 905 740788 1406 1422 114474 114490AGTGTAGGGTTAATGTT 44 906 740789 1409 1425 114477 114493CCGAGTGTAGGGTTAAT 70 907 740790 1412 1428 114480 114496ATTCCGAGTGTAGGGTT 76 908 740791 1415 1431 114483 114499GGAATTCCGAGTGTAGG 22 909 740792 1418 1434 114486 114502CAGGGAATTCCGAGTGT 75 910 740793 1422 1438 114490 114506GCTTCAGGGAATTCCGA 68 911 740794 1425 1441 114493 114509GTTGCTTCAGGGAATTC 84 912 740795 1428 1444 114496 114512AGTGTTGCTTCAGGGAA 76 913 740796 1431 1447 114499 114515GGCAGTGTTGCTTCAGG 82 914 740797 1434 1450 114502 114518TCTGGCAGTGTTGCTTC 55 915 740798 1437 1453 114505 114521ACTTCTGGCAGTGTTGC 63 916 740799 1440 1456 114508 114524CACACTTCTGGCAGTGT 19 917 740800 1444 1460 114512 114528AAAACACACTTCTGGCA 50 918 740801 1447 1463 114515 114531ACCAAAACACACTTCTG 87 919 740802 1450 1466 114518 114534CATACCAAAACACACTT 87 920 740803 1453 1469 114521 114537GTGCATACCAAAACACA 31 921 740804 1456 1472 114524 114540CCAGTGCATACCAAAAC 77 922 740805 1459 1475 114527 114543GAACCAGTGCATACCAA 67 923 740806 1462 1478 114530 114546AAGGAACCAGTGCATAC 69 924 740807 1466 1482 114534 114550ACTTAAGGAACCAGTGC 49 925 740808 1469 1485 114537 114553GCCACTTAAGGAACCAG 82 926 740809 1472 1488 114540 114556ACAGCCACTTAAGGAAC 64 927 740810 1475 1491 114543 114559ATCACAGCCACTTAAGG 28 928 740811 1478 1494 114546 114562TTAATCACAGCCACTTA 62 929 740812 1481 1497 114549 114565TAATTAATCACAGCCAC 67 930 740813 1484 1500 114552 114568CAATAATTAATCACAGC 74 931 740814 1488 1504 114556 114572CTTTCAATAATTAATCA 22 932 740815 1492 1508 114560 114576CCCACTTTCAATAATTA 20 933 740816 1521 1537 114589 114605TCTACAATAGTAGTTGG 23 934 740817 1524 1540 114592 114608CACTCTACAATAGTAGT 37 935 740818 1527 1543 114595 114611GACCACTCTACAATAGT 62 936 740819 1530 1546 114598 114614ATAGACCACTCTACAAT 55 937 740820 1533 1549 114601 114617GAAATAGACCACTCTAC 50 938 740821 1536 1552 114604 114620GGAGAAATAGACCACTC 64 939 740822 1545 1561 114613 114629GGATTGAAGGGAGAAAT 49 940 740823 1548 1564 114616 114632ACAGGATTGAAGGGAGA 71 941 740824 1551 1567 114619 114635TTGACAGGATTGAAGGG 57 942 740825 1554 1570 114622 114638ACATTGACAGGATTGAA 58 943 740826 1557 1573 114625 114641CAAACATTGACAGGATT 62 944 740827 1580 1596 114648 114664ACAGTTCCCCAAAATAC 50 945 740828 1583 1599 114651 114667ACAACAGTTCCCCAAAA  6 946 740829 1586 1602 114654 114670CAAACAACAGTTCCCCA 43 947 740830 1589 1605 114657 114673CATCAAACAACAGTTCC 48 948 740831 1592 1608 114660 114676ACACATCAAACAACAGT 68 949 740832 1595 1611 114663 114679CATACACATCAAACAAC 24 950 740833 1627 1643 114695 114711GCTCAATTAAAAATGTA 31 951 740834 1630 1646 114698 114714AAGGCTCAATTAAAAAT 26 952 740835 1637 1653 114705 114721TTAATAAAAGGCTCAAT 28 953 740836 1640 1656 114708 114724ATGTTAATAAAAGGCTC 57 954 740837 1647 1663 114715 114731ACAATATATGTTAATAA  3 955 740838 1661 1677 114729 114745TCGAGACAAAAATAACA 29 956 740839 1664 1680 114732 114748ATTTCGAGACAAAAATA 33 957 740840 1667 1683 114735 114751ATTATTTCGAGACAAAA 34 958 740841 1670 1686 114738 114754AAAATTATTTCGAGACA 47 959 740842 1673 1689 114741 114757TAAAAAATTATTTCGAG 11 960 740843 1685 1701 114753 114769ATAGATTTTAACTAAAA  0 961 740844 1706 1722 114774 114790TTCACACCAATATCAGA 64 962 740845 1709 1725 114777 114793GCATTCACACCAATATC 55 963 740846 1712 1728 114780 114796ACAGCATTCACACCAAT 73 964 740847 1715 1731 114783 114799GGTACAGCATTCACACC 46 965 740848 1718 1734 114786 114802AAAGGTACAGCATTCAC 65 966 740849 1721 1737 114789 114805CAGAAAGGTACAGCATT 56 967 740850 1724 1740 114792 114808TGTCAGAAAGGTACAGC 49 968 740851 1728 1744 114796 114812TTATTGTCAGAAAGGTA 79 969 740852 1731 1747 114799 114815TATTTATTGTCAGAAAG 52 970 740853 1734 1750 114802 114818TATTATTTATTGTCAGA 79 971 740854 1737 1753 114805 114821GAATATTATTTATTGTC 54 972 740855 1741 1757 114809 114825GGTCGAATATTATTTAT 51 973

TABLE 15Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 740410  240  256   4691   4707GAATTCCTTTACACCAC 91   33 740432  270  286   4721   4737AGTCCTTTCATGAATAC 79  591 740856 1745 1761 114813 114829TCATGGTCGAATATTAT 31  974 740857 1748 1764 114816 114832TATTCATGGTCGAATAT 24  975 740858 1751 1767 114819 114835TTTTATTCATGGTCGAA 62  976 740859 1754 1770 114822 114838TTTTTTTATTCATGGTC 77  977 740860 1771 1787 114839 114855GGAACCCACTTTTTTTT 14  978 740861 1774 1790 114842 114858CCGGGAACCCACTTTTT 25  979 740862 1777 1793 114845 114861TTCCCGGGAACCCACTT 20  980 740863 1780 1796 114848 114864TAGTTCCCGGGAACCCA 25  981 740864 1784 1800 114852 114868TGCTTAGTTCCCGGGAA 25  982 740865 1787 1803 114855 114871CACTGCTTAGTTCCCGG 51  983 740866 1790 1806 114858 114874CTACACTGCTTAGTTCC 76  984 740867 1793 1809 114861 114877CTTCTACACTGCTTAGT 37  985 740868 1796 1812 114864 114880CATCTTCTACACTGCTT 54  986 740869 1799 1815 114867 114883AATCATCTTCTACACTG 38  987 740870 1802 1818 114870 114886CAAAATCATCTTCTACA 17  988 740871 1806 1822 114874 114890TAGTCAAAATCATCTTC 40  989 740872 1809 1825 114877 114893GTGTAGTCAAAATCATC 58  990 740873 1812 1828 114880 114896AGGGTGTAGTCAAAATC 61  991 740874 1815 1831 114883 114899AGGAGGGTGTAGTCAAA 43  992 740875 1818 1834 114886 114902CTAAGGAGGGTGTAGTC 41  993 740876 1821 1837 114889 114905TCTCTAAGGAGGGTGTA 43  994 740877 1824 1840 114892 114908GGCTCTCTAAGGAGGGT 38  995 740878 1828 1844 114896 114912TTATGGCTCTCTAAGGA 37  996 740879 1831 1847 114899 114915GTCTTATGGCTCTCTAA 66  997 740880 1834 1850 114902 114918TGTGTCTTATGGCTCTC 72  998 740881 1837 1853 114905 114921TAATGTGTCTTATGGCT 67  999 740882 1840 1856 114908 114924TGCTAATGTGTCTTATG 59 1000 740883 1843 1859 114911 114927ATGTGCTAATGTGTCTT 66 1001 740884 1846 1862 114914 114930AATATGTGCTAATGTGT 74 1002 740885 1850 1866 114918 114934TGCTAATATGTGCTAAT 33 1003 740886 1853 1869 114921 114937ATGTGCTAATATGTGCT 34 1004 740887 1856 1872 114924 114940TGAATGTGCTAATATGT 52 1005 740888 1859 1875 114927 114943CCTTGAATGTGCTAATA 59 1006 740889 1862 1878 114930 114946GAGCCTTGAATGTGCTA 28 1007 740890 1865 1881 114933 114949TCAGAGCCTTGAATGTG 52 1008 740891 1868 1884 114936 114952CTCTCAGAGCCTTGAAT 48 1009 740892 1870 1886 114938 114954TTCTCTCAGAGCCTTGA 74 1010 740893 1871 1887 114939 114955ATTCTCTCAGAGCCTTG 83  444 740894 1872 1888 114940 114956CATTCTCTCAGAGCCTT 80 1011 740895 1874 1890 114942 114958CACATTCTCTCAGAGCC 57 1012 740896 1875 1891 114943 114959CCACATTCTCTCAGAGC 57 1013 740897 1995 2011 115063 115079TAAAGGCATTTCCTGTA 50 1014 740898 2081 2097 115149 115165GGCAACATTTAAAGGAG 46 1015 740899 2251 2267 115319 115335GAAATGACTATGCCCCA 61 1016 740900 2312 2328 115380 115396AAACTGCTAGCATGTCT 62 1017 740901 2437 2453 115505 115521TGTAGTAGTCTCTCTTC 77 1018 740902 2841 2857 115909 115925TGTTTAAGTGTTTGGTC 79 1019 740903 2939 2955 116007 116023GGACTGGATTGATCCTC 48 1020 740904 3158 3174 116226 116242ACATACTGGATAAGCCA 83 1021 740905 N/A N/A   2087   2103 CCTGGATCACACCAGAA28 1022 740906 N/A N/A   2090   2106 GTTCCTGGATCACACCA 45 1023 740907N/A N/A   2093   2109 GCTGTTCCTGGATCACA 41 1024 740908 N/A N/A   2096  2112 ACAGCTGTTCCTGGATC  2 1025 740909 N/A N/A   2099   2115AAGACAGCTGTTCCTGG 19 1026 740910 N/A N/A   2102   2118 TGGAAGACAGCTGTTCC 7 1027 740911 N/A N/A   2105   2121 AGCTGGAAGACAGCTGT 13 1028 740912N/A N/A   2108   2124 CAGAGCTGGAAGACAGC 26 1029 740913 N/A N/A   2113  2129 TCTTTCAGAGCTGGAAG 16 1030

TABLE 16Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 97  33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC 92 591 740933 N/A N/A  3624  3640 CCTGAGTCCCTCTGCTT 27 1031 740934 N/A N/A 3849  3865 TAATCTCTCAGCCCTTG 55 1032 740935 N/A N/A  4074  4090CCACCCTAGCGGACCCC 35 1033 740936 N/A N/A  4299  4315 CCAGAAGAAGGCTAACA35 1034 740937 N/A N/A  4524  4540 GGCATTAAAAATTTAGC 68 1035 740938 N/AN/A  4684  4700 TTTACACCACACTGGAA 65 1036 740939 N/A N/A  4685  4701CTTTACACCACACTGGA 82 1037 740940 N/A N/A  4686  4702 CCTTTACACCACACTGG68 1038 740941 N/A N/A  4821  4837 ACCTACATAGAGAACAC 83 1039 740942 N/AN/A  5046  5062 GGCAAAATTAAAAATCT 38 1040 740943 N/A N/A  5275  5291TTATACACATCACAGGG 59 1041 740944 N/A N/A  5500  5516 AAGGTGGAACTTTAGGA73 1042 740945 N/A N/A  5725  5741 GACTCTTACTGCTATAG 47 1043 740946 N/AN/A  5984  6000 TGATAGCATCACTGCAG 13 1044 740947 N/A N/A  6209  6225AACTCATCAATTTTTTC 67 1045 740948 N/A N/A  6439  6455 GTAACCAATAAAAAATT24 1046 740949 N/A N/A  6715  6731 GTTGTTTGTAGACACAG 54 1047 740950 N/AN/A  6940  6956 TGTTTATGACTACCTTC 62 1048 740951 N/A N/A  7165  7181ATTTTTTACTAATCAGG 38 1049 740952 N/A N/A  7615  7631 GTCATTTGAAGAAATTT60 1050 740953 N/A N/A  7840  7856 GTGCATGTTATGTTGAC 36 1051 740954 N/AN/A  8065  8081 TTATGAGTAATCTGTAA 30 1052 740955 N/A N/A  8290  8306GCCACTAAACCACACCA 65 1053 740956 N/A N/A  8544  8560 GGGATGATGAGATCAGG40 1054 740957 N/A N/A  8769  8785 TTTTAGCTGCCCTTGCC 25 1055 740958 N/AN/A  8995  9011 TTATCTCACATATATGT 30 1056 740959 N/A N/A  9240  9256ACACCACTCCATTGCAG 46 1057 740960 N/A N/A  9465  9481 GGAGTGGACATGTTTTT43 1058 740961 N/A N/A  9691   9707 CAACACAGTGGCTCTTG 24 1059 740962 N/AN/A  9920   9936 GAATGATAAATGTTTCA 32 1060 740963 N/A N/A 10146 10162AGATAGAAGTAGAGAGT 14 1061 740964 N/A N/A 10371 10387 TTGTTTGTGCTGGAACT16 1062 740965 N/A N/A 10596 10612 CATAACAGATGTGAAGC 45 1063 740966 N/AN/A 10821 10837 TGCAGCAGTGACAACAT 73 1064 740967 N/A N/A 11046 11062TTTACAGAATTATCATA 37 1065 740968 N/A N/A 11271 11287 CATTACACATGTAATAA 6 1066 740969 N/A N/A 11729 11745 CATTATGTAAAAAAAAC  0 1067 740970 N/AN/A 11954 11970 TACGATTTTAGCACAAA 68 1068 740971 N/A N/A 12182 12198CCTACAAAAACAAATTC  0 1069 740972 N/A N/A 12192 12208 GGTTTTGGAGCCTACAA83 1070 740973 N/A N/A 12421 12437 GCAAGTATATTTTTTAT 62 1071 740974 N/AN/A 12646 12662 CCTGAAATGCACTCTGA 54 1072 740975 N/A N/A 12871 12887CTCATCTTCCTCAACAT 56 1073 740976 N/A N/A 13098 13114 TCCATTTTAGAAGTCAG87 1074 740977 N/A N/A 13331 13347 TAACACTTATAAAATAC 44 1075 740978 N/AN/A 13556 13572 GAGGTCCCTAGAAGGCA 38 1076 740979 N/A N/A 13781 13797TCTCCATTAGATCATCA 43 1077 740980 N/A N/A 14011 14027 GAGAAAATAAAGTATAC41 1078 740981 N/A N/A 14236 14252 TGGTCCATGGGTGCAAT 52 1079 740982 N/AN/A 14461 14477 ATATGCAAATTATTCTC 40 1080 740983 N/A N/A 14686 14702TTCCCAGCCCAAGTTTA  1 1081 740984 N/A N/A 14911 14927 AATAGGTAACTTTATAT19 1082 740985 N/A N/A 15136 15152 TAATATATGGTTTTGAA 28 1083 740986 N/AN/A 15365 15381 GGATTCTGCTTTATTTT 51 1084 740987 N/A N/A 15590 15606CGACACATTTAAAAACA 36 1085 740988 N/A N/A 15815 15831 AAAGCGAGATTAAAAAT 0 1086 740989 N/A N/A 16040 16056 GGATATGGCTGATGTCT 13 1087 740990 N/AN/A 16265 16281 CCAATATTTAAATGGTG 34 1088 740991 N/A N/A 16591 16607GCCAATATTTACTTATT 61 1089 740992 N/A N/A 16818 16834 TCATGTGGAATCTAAAG 6 1090 740993 N/A N/A 17043 17059 AGTATGAAAATGAAGAG 38 1091 740994 N/AN/A 17501 17517 ATTCTTGTTGTTCAGGC 73 1092 740995 N/A N/A 17726 17742GGAATGTAAAGCCATGA 78 1093 740996 N/A N/A 17951 17967 ATTAAAGGGTGGTAGAA26 1094 740997 N/A N/A 18176 18192 AATGAACCGTAATCTCA 87 1095 740998 N/AN/A 18296 18312 CTGGGTTAATGCCTGAA 60 1096 740999 N/A N/A 18297 18313TCTGGGTTAATGCCTGA 67  163 741000 N/A N/A 18298 18314 ATCTGGGTTAATGCCTG86 1097 741001 N/A N/A 18401 18417 GAAATGTCACTGTTCCT 97 1098 741002 N/AN/A 18626 18642 GGTTGGTATGTATTTTA 92 1099 741003 N/A N/A 18851 18867CAAGGAGGTCATTGTGG 75 1100 741004 N/A N/A 19183 19199 CTTTGGCAAAGAAAGGA45 1101 741005 N/A N/A 19408 19424 AAATGAAAGTTGTTGTG 85 1102 741006 N/AN/A 19633 19649 GATATTTTTGTTCTGCC 95 1103 741007 N/A N/A 19868 19884GCTATAAATAGAATTAA 42 1104 741008 N/A N/A 20099 20115 TAGATTTCTGTTTCCTC94 1105 741009 N/A N/A 20324 20340 AATGCAGGTGAATAAAA 81 1106

TABLE 17Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 92   33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC87  591 741010 N/A N/A 20549 20565 CAATTTCTAGGTTCTAT 86 1107 741011 N/AN/A 20559 20575 AACTATGCTGCAATTTC 74 1108 741012 N/A N/A 20561 20577ACAACTATGCTGCAATT 86 1109 741013 N/A N/A 20562 20578 CACAACTATGCTGCAAT88  314 741014 N/A N/A 20565 20581 TTCCACAACTATGCTGC 90 1110 741015 N/AN/A 20774 20790 GACCACAATTGCAGACA 86 1111 741016 N/A N/A 20985 21001GTGTGAGCAAACATTCT 94 1112 741017 N/A N/A 27412 27428 CAGTGTGAGCAAACATT90 1113 N/A N/A 20987 21003 741018 N/A N/A 27413 27429 ACAGTGTGAGCAAACAT85  468 N/A N/A 20988 21004 741019 N/A N/A 27414 27430 CACAGTGTGAGCAAACA91 1114 N/A N/A 20989 21005 741020 N/A N/A 20991 21007 GGCACAGTGTGAGCAAA89 1115 741021 N/A N/A 20999 21015 AAGTTTCTGGCACAGTG 89 1116 741022 N/AN/A 21224 21240 GTTCAGAATTATGTCAT 95 1117 741023 N/A N/A 21449 21465TCTTATGTGCACATGAG 63 1118 741024 N/A N/A 21674 21690 CATAGTAGCATTACAGA83 1119 741025 N/A N/A 21899 21915 TCAGGCAGTGGCTTCAC 66 1120 741026 N/AN/A 22129 22145 TAAAAAAAGTTGTTCAT 32 1121 741027 N/A N/A 22360 22376CACTCAAGTGTTTAAAA 87 1122 741028 N/A N/A 22454 22470 TGTGACCTGTGCTTGTT91 1123 741029 N/A N/A 22456 22472 CCTGTGACCTGTGCTTG 87 1124 741030 N/AN/A 22457 22473 GCCTGTGACCTGTGCTT 86   88 741031 N/A N/A 22458 22474TGCCTGTGACCTGTGCT 87 1125 741032 N/A N/A 22460 22476 GTTGCCTGTGACCTGTG82 1126 741033 N/A N/A 22599 22615 TATTAGACACTTAAGGG 82 1127 741034 N/AN/A 22831 22847 TCAATCTTAAATTTTTC 85 1128 741035 N/A N/A 23056 23072GTACTTTCCCACCTAGA 88 1129 741036 N/A N/A 23281 23297 TCTCAGAGACCACAGCT88 1130 741037 N/A N/A 23285 23301 TTGTTCTCAGAGACCAC 92 1131 741038 N/AN/A 23286 23302 ATTGTTCTCAGAGACCA 86  164 741039 N/A N/A 23287 23303TATTGTTCTCAGAGACC 94 1132 741040 N/A N/A 23289 23305 CATATTGTTCTCAGAGA89 1133 741041 N/A N/A 23506 23522 ACTATTAACCACTGATC 84 1134 741042 N/AN/A 23731 23747 GTTGCAGTCCACAGAAT 79 1135 741043 N/A N/A 23956 23972TAAAGATAAGTATCTCA 91 1136 741044 N/A N/A 24181 24197 AAAACAAACCTAAGTCA43 1137 741045 N/A N/A 24406 24422 AAAAGCTAACAGCCTAT 73 1138 741046 N/AN/A 24631 24647 TTAAATTGATGAGATGT 88 1139 741047 N/A N/A 24856 24872GTATTCTTTGCATTAGT 89 1140 741048 N/A N/A 25081 25097 TAAAAGTGTACATTATT77 1141 741049 N/A N/A 25306 25322 CTCAAGGCAAAGCTGTA 88 1142 741050 N/AN/A 25531 25547 TGCCACTATAAGCAGTC 94 1143 741051 N/A N/A 25756 25772TTCAAGCCCATGCCCTC 84 1144 741052 N/A N/A 25801 25817 ATCCAGTAGAGTGAGAG79 1145 741053 N/A N/A 25803 25819 TCATCCAGTAGAGTGAG 89 1146 741054 N/AN/A 25804 25820 ATCATCCAGTAGAGTGA 85  315 741055 N/A N/A 25807 25823GACATCATCCAGTAGAG 92 1147 741056 N/A N/A 25923 25939 TGAATACATTGTCTTAA81 1148 741057 N/A N/A 25925 25941 ATTGAATACATTGTCTT 90 1149 741058 N/AN/A 25926 25942 AATTGAATACATTGTCT 94  392 741059 N/A N/A 25927 25943TAATTGAATACATTGTC 84 1150 741060 N/A N/A 25929 25945 CATAATTGAATACATTG80 1151 741061 N/A N/A 25981 25997 TGAGTAGCTATGGTTTA 91 1152 741062 N/AN/A 26202 26218 TCTTTGTGTTATACAAT 58 1153 741063 N/A N/A 26204 26220CCTCTTTGTGTTATACA 83 1154 741064 N/A N/A 26205 26221 CCCTCTTTGTGTTATAC87  469 741065 N/A N/A 26206 26222 TCCCTCTTTGTGTTATA 77 1155 741066 N/AN/A 26208 26224 TTTCCCTCTTTGTGTTA 78 1156 741067 N/A N/A 26431 26447TACATACAATATTAAGG 78 1157 741068 N/A N/A 26656 26672 AAAAGAATGGATTCTGA75 1158 741069 N/A N/A 26881 26897 AAGGAAAAACTCTGCCC 73 1159 741070 N/AN/A 27106 27122 TCACCCCAAGGCATTTG 56 1160 741071 N/A N/A 27331 27347ACACCCTGATTCCCAAG 82 1161 741072 N/A N/A 27410 27426 GTGTGAGCAAACATTCA89 1162 741073 N/A N/A 27416 27432 GTCACAGTGTGAGCAAA 91 1163 741074 N/AN/A 27556 27572 GGGAAGTATTAGTGGAA 86 1164 741075 N/A N/A 27782 27798GCTGAAAATATGAAACA 76 1165 741076 N/A N/A 28007 28023 ACTTCTAGCACTATTTT71 1166 741077 N/A N/A 28232 28248 TTGTGCATTTATTCCAC 93 1167 741078 N/AN/A 28457 28473 GACTGTAATCTAGGACC 90 1168 741079 N/A N/A 28682 28698TGACTTTTGAATCAGTC 59 1169 741080 N/A N/A 29010 29026 GAGCGATTCTCCTGGTT76 1170 741081 N/A N/A 29235 29251 CACAGTCCATAATATTG 79 1171 741082 N/AN/A 29460 29476 TTTTTGTTAATAGTTCT 80 1172 741083 N/A N/A 29685 29701GCTTTCTCAGAGCCCAA 89 1173 741084 N/A N/A 29912 29928 ATCTCTCTACCATGTGA79 1174 741085 N/A N/A 30137 30153 GTGGATAAAGTACATTA 77 1175 741086 N/AN/A 30362 30378 AAATGGTATTCAGAGAT 76 1176

TABLE 18Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 91   33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC94  591 741087 N/A N/A 30587 30603 TTTTCTCCTAAAGCCTT 85 1177 741088 N/AN/A 31037 31053 CAGATTTCCAGCACACT 81 1178 741089 N/A N/A 31262 31278CCTTCTTAGTGGTAAGA 64 1179 741090 N/A N/A 31487 31503 AATTACAGTGTAGGTAA52 1180 741091 N/A N/A 31712 31728 ATAAGAGGTCACTGGAT 91 1181 741092 N/AN/A 31937 31953 AAGGAAACAGTCTACAT 85 1182 741093 N/A N/A 32162 32178CTATCATGATAAGTATA 84 1183 741094 N/A N/A 32387 32403 TGTGGTTCTGCCCATCT88 1184 741095 N/A N/A 32624 32640 GCCTAAACATTTTACTT 34 1185 741096 N/AN/A 32858 32874 GAAGTTTCTGAAGAAAT 89 1186 741097 N/A N/A 33083 33099TTTTCAGTAGATTTGAC 86 1187 741098 N/A N/A 33308 33324 GCTATGACCCTCAAGCC68 1188 741099 N/A N/A 33533 33549 AATAGAGCAAAATTTCG 87 1189 741100 N/AN/A 33762 33778 ATAATCAAACAAAAGGG 73 1190 741101 N/A N/A 33987 34003AAAGTTCAATGCTGTGT 94 1191 741102 N/A N/A 34212 34228 GAAATGGGCATGTAAAC72 1192 741103 N/A N/A 34443 34459 CAAAATACAATGTTCAA 40 1193 741104 N/AN/A 34668 34684 ATTCTTCTATCCTAGAA 12 1194 741105 N/A N/A 34893 34909ATTATCATGGTTGCCCA 91 1195 741106 N/A N/A 35118 35134 ATGAGATCTTTTTGCAT87 1196 741107 N/A N/A 35343 35359 AAGCAAGTTGTCCATGG 90 1197 741108 N/AN/A 35568 35584 TGTTGGAGTTTACAATT 76 1198 741109 N/A N/A 35793 35809CTCACTAGCCCTGTGAC 14 1199 741110 N/A N/A 36018 36034 TCTCTTTCATGGGTATT92 1200 741111 N/A N/A 36252 36268 GTCATTTTAATAAGTGT 92 1201 741112 N/AN/A 36484 36500 CAATTAAATAAACCTCT 65 1202 741113 N/A N/A 36790 36806TATGGTGATATGGTTAG 91 1203 741114 N/A N/A 37018 37034 CCATGTGTTTTTGTGGC84 1204 741115 N/A N/A 37243 37259 CAAAGGTATAAGGTCAT 94 1205 741116 N/AN/A 37468 37484 AGCTTGTATTTTTGAAA 86 1206 741117 N/A N/A 37788 37804CGCATCTGTCTTTCTTT 78 1207 741118 N/A N/A 38013 38029 TAGGACAGGTGAAATAA72 1208 741119 N/A N/A 38238 38254 AGTTATTAGAATAACAC  0 1209 741120 N/AN/A 38464 38480 AATAAAATGTCTTAATC 25 1210 741121 N/A N/A 38691 38707ACTCAAAAAAGAAGAAT 44 1211 741122 N/A N/A 38916 38932 GTTTTCTCTGTATTGGC93 1212 741123 N/A N/A 39141 39157 TGGCCTAGTGGTTATAA 19 1213 741124 N/AN/A 39366 39382 CACAAAGAGGAAACAGG 80 1214 741125 N/A N/A 39591 39607ACATTTTTTAACTGGAT 92 1215 741126 N/A N/A 39816 39832 AGGCTAAATTTTAATAA 6 1216 741127 N/A N/A 40041 40057 TAGCCTTTCATAGTACG 90 1217 741128 N/AN/A 40266 40282 AAGAGGAAAAGCTTGGA 43 1218 741129 N/A N/A 40491 40507AAAAATTCTGGTGCCAA 94 1219 741130 N/A N/A 40716 40732 AAGCTAAACTACCGCTG58 1220 741131 N/A N/A 40941 40957 GAATTTCCTGGATGCTC 92 1221 741132 N/AN/A 41130 41146 AGATTCCAGCAGAGATT 74 1222 741133 N/A N/A 41132 41148ACAGATTCCAGCAGAGA 89 1223 741134 N/A N/A 41133 41149 AACAGATTCCAGCAGAG87   90 741135 N/A N/A 41134 41150 GAACAGATTCCAGCAGA 85 1224 741136 N/AN/A 41136 41152 GTGAACAGATTCCAGCA 86 1225 741137 N/A N/A 41166 41182ATCTGTAAGAAGTTTAG 52 1226 741138 N/A N/A 41391 41407 TGAGAAATTTTATGGGT86 1227 741139 N/A N/A 41620 41636 TCATTCAAAACCATCCT 78 1228 741140 N/AN/A 41845 41861 GATCACACTGCTTATAG 84 1229 741141 N/A N/A 42070 42086CAAGTTGATGGCATATA 89 1230 741142 N/A N/A 42295 42311 GTGTACCAACCTCAAGT71 1231 741143 N/A N/A 42532 42548 TAAGTAAATACCTAGGG 83 1232 741144 N/AN/A 42757 42773 GATTTGTGCCTGGCATC 91 1233 741145 N/A N/A 42835 42851TGCCTCTACCTCCAGCA 89 1234 741146 N/A N/A 42837 42853 GATGCCTCTACCTCCAG87 1235 741147 N/A N/A 42838 42854 TGATGCCTCTACCTCCA 85  166 741148 N/AN/A 42839 42855 CTGATGCCTCTACCTCC 87 1236 741149 N/A N/A 42982 42998TATCACAACTACATTGT 40 1237 741150 N/A N/A 43208 43224 GGCCTCCTGCTGCAGCA31 1238 741151 N/A N/A 43440 43456 GCACTCATTTTAAATGT 72 1239 741152 N/AN/A 43665 43681 TGGTAACTTAGGACAAG 93 1240 741153 N/A N/A 43818 43834TTCTCTGGACCTCTTAA 67 1241 741154 N/A N/A 43820 43836 ACTTCTCTGGACCTCTT85 1242 741155 N/A N/A 43821 43837 TACTTCTCTGGACCTCT 92  242 741156 N/AN/A 43822 43838 TTACTTCTCTGGACCTC 90 1243 741157 N/A N/A 43890 43906TCAATACAACTTAATTC 48 1244 741158 N/A N/A 44376 44392 TTGGGCTGGAAGCAGTG43 1245 741159 N/A N/A 44601 44617 AAGATATGCAGAGGGTT 92 1246 741160 N/AN/A 44828 44844 TGGTCTAACTGTGTTGC 85 1247 741161 N/A N/A 45053 45069GTTTATGGACTTTTTAA 87 1248 741162 N/A N/A 45278 45294 TTTTGTACTTTATGGAA89 1249 741163 N/A N/A 45503 45519 ACTTCTCCTTCAATTAA 72 1250

TABLE 19Percent reduction of human SNCA mRNA with 4-9-4 MOE and cEt gapmerswith mixed internucleoside linkages SEQ ID SEQ ID SEQ ID SEQ ID CompoundNo: 1 No: 1 No: 2 No: 2 % SEQ No start stop start stopSequence (5′ to 3′) Reduction ID NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 90   33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC90  591 741241 N/A N/A 56397 56413 AAAATTTTTGCACACTT 94 1251 741242 N/AN/A 56622 56638 GCCAAATCAATGGATGA 90 1252 741243 N/A N/A 56847 56863AGTGACCAAGAGAATGA 53 1253 741244 N/A N/A 57072 57088 TTTTAAAACACTGGCCT41 1254 741245 N/A N/A 57297 57313 TAGGATTAAACAGTCCA 48 1255 741246 N/AN/A 57522 57538 TTATCTGTTGCTATGTG 91 1256 741247 N/A N/A 57747 57763AAGAAGGAGAATAGCAG 86 1257 741248 N/A N/A 57981 57997 CGGGCAAACATGTTTTG47 1258 741249 N/A N/A 58206 58222 ATGACCTACATGCTAAA 73 1259 741250 N/AN/A 58431 58447 AGAAGCAAAATGTCAGT 88 1260 741251 N/A N/A 58656 58672CCTAACAGCTTTACTTT 50 1261 741252 N/A N/A 58881 58897 CTTTCACACATCTCTAA64 1262 741253 N/A N/A 58991 59007 TTTCATTAATCTGTGAA 34 1263 741254 N/AN/A 58992 59008 ATTTCATTAATCTGTGA 86  169 741255 N/A N/A 58993 59009TATTTCATTAATCTGTG 93 1264 741256 N/A N/A 58995 59011 TATATTTCATTAATCTG87 1265 741257 N/A N/A 59106 59122 CCTTACACAAAATATAA 38 1266 741258 N/AN/A 59354 59370 ACACCAATATATTATTT 67 1267 741259 N/A N/A 59594 59610TAAAGGATGCAAAGGCA 55 1268 741260 N/A N/A 59948 59964 TTCCAGCGATCCCACTC80 1269 741261 N/A N/A 60173 60189 CTCAACATCTTTAATGA 35 1270 741262 N/AN/A 60421 60437 GGGACCTAAAACTATAA 25 1271 741263 N/A N/A 60758 60774AGCAGAATAGAAAATCC 49 1272 741264 N/A N/A 60983 60999 TTCAATGCGACTCCCAT81 1273 741265 N/A N/A 61216 61232 CAACAAAACTGAGAATC 24 1274 741266 N/AN/A 61474 61490 AATGCCTGCTTTCACCA 76 1275 741267 N/A N/A 61699 61715TATAAGCAGGAGTAAAA 27 1276 741268 N/A N/A 61969 61985 GTTCCAAAAGATAGAGA55 1277 741269 N/A N/A 62200 62216 CGTACACAAACTAGAAA 33 1278 741270 N/AN/A 62492 62508 TACTGTTGCATTCCAGC 70 1279 741271 N/A N/A 62729 62745TCTTAGTGTGGTGGCTC 78 1280 741272 N/A N/A 62955 62971 TCAACAATAATAATGAC60 1281 741273 N/A N/A 63197 63213 CCTTTTCATCAACACAT 71 1282 741274 N/AN/A 63422 63438 TATGCATCTAACACTTG 52 1283 741275 N/A N/A 63666 63682CCATCAACCAAGTATCT 26 1284 741276 N/A N/A 63891 63907 CTTGAAACAGTAACTTG47 1285 741277 N/A N/A 64116 64132 AACATAGCAGATTAATA 37 1286 741278 N/AN/A 64349 64365 TCATGTTATATAGTGGG 97 1287 741279 N/A N/A 64574 64590TGTAACCTAATGTAAAT 37 1288 741280 N/A N/A 64799 64815 ACAAGTATCTGTACTCA94 1289 741281 N/A N/A 65024 65040 GTCTCTGTTAATGTTGG 75 1290 741282 N/AN/A 65249 65265 GAACCAGCCTGACTTAA 74 1291 741283 N/A N/A 65474 65490TTGTATGGGTTACATAA 61 1292 741284 N/A N/A 65801 65817 CAATTAAATGCAATTCC53 1293 741285 N/A N/A 66026 66042 TGACAGAAGTGTGCATA 59 1294 741286 N/AN/A 66251 66267 CAACACATCCACATTGC 75 1295 741287 N/A N/A 66476 66492TTCACACCTCTCTCCCT 51 1296 741288 N/A N/A 66701 66717 TGCTGGTCTAAGATGCA77 1297 741289 N/A N/A 66926 66942 ATGTGTTTTGAGGAAAA 77 1298 741290 N/AN/A 67151 67167 CAGAAGTAAATGTGGAC 85 1299 741291 N/A N/A 67376 67392TGATTCTTTGGATTCAT 79 1300 741292 N/A N/A 67876 67892 CATTCTTGTTTTTATTC86 1301 741293 N/A N/A 68101 68117 AATAGTGTCCCAGTGTA 78 1302 741294 N/AN/A 68326 68342 TGAAAGCTGTTCAGTTA 74 1303 741295 N/A N/A 68551 68567CCCACATATACTACTTG 86 1304 741296 N/A N/A 68776 68792 AGAATTTCAGGAAGTTA87 1305 741297 N/A N/A 68798 68814 CAAAGTAAGAGGAGATT 62 1306 741298 N/AN/A 68800 68816 GCCAAAGTAAGAGGAGA 90 1307 741299 N/A N/A 68801 68817TGCCAAAGTAAGAGGAG 61  397 741300 N/A N/A 68804 68820 CAGTGCCAAAGTAAGAG64 1308 741301 N/A N/A 69001 69017 TGAATCCATTTGTCCAG 91 1309 741302 N/AN/A 69227 69243 CTCTAAAATACAAATGT 72 1310 741303 N/A N/A 69452 69468GAACAAAGGAATAAGTA 59 1311 741304 N/A N/A 69677 69693 CTAGATGTAGATATCAT61 1312 741305 N/A N/A 69902 69918 AAGGGAATAAATTGTAG 52 1313 741306 N/AN/A 70127 70143 CAACAGACCCTTTCAAT 60 1314 741307 N/A N/A 70352 70368GTCTTCCCACTGCCTAC 62 1315 741308 N/A N/A 70577 70593 TTTAGATATACCTCCAA94 1316 741309 N/A N/A 70880 70896 GCTTCAGTTTCTTGAGT 79 1317 741310 N/AN/A 71105 71121 CTGGTCTTTCTCACAAT N.D. 1318 741311 N/A N/A 71375 71391ATCATTCTTAACAGAAA 70 1319 741312 N/A N/A 71600 71616 GCTCTTGCTGTGCAGCC74 1320 741313 N/A N/A 71844 71860 ATTTAAAGCAGCAGTCC 50 1321 741314 N/AN/A 72076 72092 AGGTAATTCTAATTTTA 68 1322 741315 N/A N/A 72301 72317GGCAAATGACAGGGTCT 93 1323 741316 N/A N/A 72632 72648 TCTCAACTGCCTGAGTA22 1324 741317 N/A N/A 72857 72873 CATGTCAGCTTTTTAGT 69 1325

TABLE 20 Percent reduction of human SNCA mRNA with 4-9-4 MOEand cEt gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 Sequence % ID No start stopstart stop (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 95   33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC94  591 741318 N/A N/A 73090 73106 ATACTCAGATATTTAAA 64 1326 741319 N/AN/A 73188 73204 ACTTTCTGTGTGGTATG 91 1327 741320 N/A N/A 73190 73206AGACTTTCTGTGTGGTA 95 1328 741321 N/A N/A 73191 73207 CAGACTTTCTGTGTGGT93  170 741322 N/A N/A 73192 73208 ACAGACTTTCTGTGTGG 86 1329 741323 N/AN/A 73194 73210 AGACAGACTTTCTGTGT 46 1330 741324 N/A N/A 73315 73331GTTGAGAATTTTTCATT 66 1331 741325 N/A N/A 73540 73556 AGTTATGGAGCATCTTT89 1332 741326 N/A N/A 73765 73781 GACTGAGTTTTTTATTC 74 1333 741327 N/AN/A 73990 74006 TCCTGAATTAAAAATTT 13 1334 741328 N/A N/A 74215 74231GCTAAGCACAAACAATT 62 1335 741329 N/A N/A 74292 74308 GAACTCTGTAGTCAGAA92 1336 741330 N/A N/A 74294 74310 TAGAACTCTGTAGTCAG 93 1337 741331 N/AN/A 74295 74311 ATAGAACTCTGTAGTCA 95  398 741332 N/A N/A 74296 74312AATAGAACTCTGTAGTC 92 1338 741333 N/A N/A 74298 74314 TGAATAGAACTCTGTAG85 1339 741334 N/A N/A 74440 74456 ACACAGAGCACTTCTTA 70 1340 741335 N/AN/A 74665 74681 GGAGTTACAGAGTTGCC 91 1341 741336 N/A N/A 74890 74906TATCAGTCTATTAAGAA 82 1342 741337 N/A N/A 75115 75131 AAGTTTCTCAGAGCCTG82 1343 741338 N/A N/A 75340 75356 AATACAGAAGTCTATTC 68 1344 741339 N/AN/A 75573 75589 CATTGAATAAAAATTTG 18 1345 741340 N/A N/A 75945 75961CAGGTATAAAATTTTTT 45 1346 741341 N/A N/A 76170 76186 GGTGTTAATCACTTGAA86 1347 741342 N/A N/A 76398 76414 TCTTGAAGCTAGTTGGG 91 1348 741343 N/AN/A 76623 76639 AGGGCAACTAACCAACA 75 1349 741344 N/A N/A 76848 76864GTGGATACTTAGTATCA 69 1350 741345 N/A N/A 77073 77089 CTCTCTCAGTTGTAGGT67 1351 741346 N/A N/A 77298 77314 AAAGTATGCTGTGTTCT 92 1352 741347 N/AN/A 77523 77539 GTACCCGGCACTTTTCC 53 1353 741348 N/A N/A 77663 77679TCTAGAAAAGCTCTCTT 57 1354 741349 N/A N/A 77665 77681 ACTCTAGAAAAGCTCTC81 1355 741350 N/A N/A 77666 77682 GACTCTAGAAAAGCTCT 91  247 741351 N/AN/A 77667 77683 AGACTCTAGAAAAGCTC 84 1356 741352 N/A N/A 77748 77764TGGCACCCAGGAGTAAG 65 1357 741353 N/A N/A 77973 77989 CATACACAAAATCCCCT83 1358 741354 N/A N/A 78198 78214 CACATGAAGCCAGGGAC 77 1359 741355 N/AN/A 78423 78439 GCAGGCCCTAAACTGTG 39 1360 741356 N/A N/A 78648 78664AAATTTATCTATCATGC 93 1361 741357 N/A N/A 78873 78889 GCTAAACACTTTATCAA75 1362 741358 N/A N/A 79098 79114 ACTTCATTCTTTCTGTT 87 1363 741359 N/AN/A 79323 79339 CAATTAAAAGATTACTT  0 1364 741360 N/A N/A 79548 79564ACATTGTACAGTTAATT 77 1365 741361 N/A N/A 79773 79789 TACAAACCTTACTATGC51 1366 741362 N/A N/A 79998 80014 AACAGACTTAAACAAAC 88 1367 741363 N/AN/A 80223 80239 CTCAGACATCATGTTTT 91 1368 741364 N/A N/A 80448 80464AGGCACTCACAAACATT 86 1369 741365 N/A N/A 80673 80689 TCTCGCATCCTAAATGT50 1370 741366 N/A N/A 80898 80914 TTCATATTTTATGTTAC 89 1371 741367 N/AN/A 80991 81007 TGAAATTTTCCAGCTAA 93 1372 741368 N/A N/A 80993 81009CTTGAAATTTTCCAGCT 97 1373 741369 N/A N/A 80995 81011 ATCTTGAAATTTTCCAG86 1374 741370 N/A N/A 81123 81139 CTATAATTACATTCCTA 73 1375 741371 N/AN/A 81348 81364 GCATGAACCTAGATATG 58 1376 741372 N/A N/A 81472 81488GCTGTTTGAAGTGACAA 70 1377 741373 N/A N/A 81474 81490 GAGCTGTTTGAAGTGAC90 1378 741374 N/A N/A 81475 81491 AGAGCTGTTTGAAGTGA 82  249 741375 N/AN/A 81476 81492 GAGAGCTGTTTGAAGTG 76 1379 741376 N/A N/A 81478 81494TGGAGAGCTGTTTGAAG 69 1380 741377 N/A N/A 81575 81591 CTGCCACTATTCACAAT71 1381 741378 N/A N/A 81800 81816 TTATTGCATTAATGGAA 94 1382 741379 N/AN/A 82107 82123 ATGGTGTTAGCTAGGAT 91 1383 741380 N/A N/A 82332 82348GTCTTTTTACATTATAA 93 1384 741381 N/A N/A 82557 82573 ATAACCACTATTCAATG63 1385 741382 N/A N/A 82783 82799 AAAAATCACATTTGGCA 95 1386 741383 N/AN/A 83008 83024 TTCTTTCACCTTATGAG 72 1387 741384 N/A N/A 83233 83249ATATATGTGTCAGTTCT 90 1388 741385 N/A N/A 83458 83474 GTGTCACTTTTTAAGGT14 1389 741386 N/A N/A 83528 83544 AGAACAATGTCATCTTT 94 1390 741387 N/AN/A 83530 83546 AAAGAACAATGTCATCT 88 1391 741388 N/A N/A 83531 83547GAAAGAACAATGTCATC 89   98 741389 N/A N/A 83532 83548 GGAAAGAACAATGTCAT82 1392 741390 N/A N/A 83534 83550 CAGGAAAGAACAATGTC 88 1393 741391 N/AN/A 83683 83699 CACAGGTATACACACTT 90 1394 741392 N/A N/A 83908 83924GTACAAAATCTGCATAT 81 1395 741393 N/A N/A 84133 84149 ATAGGTATTTTATGCAT88 1396 741394 N/A N/A 84616 84632 CAAATTATGCATTTGTT 66 1397

TABLE 21 Percent reduction of human SNCA mRNA with 4-9-4 MOEand cEt gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 Sequence % ID No start stopstart stop (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 95   33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC93  591 741395 N/A N/A 84845 84861 CTACAAAATTGCTAAAA  9 1398 741396 N/AN/A 85070 85086 ACAATGTTACTTTGTCC 90 1399 741397 N/A N/A 85295 85311TACAAAAATACCCCCCC 12 1400 741398 N/A N/A 85520 85536 ATTTGTAGACCTATGTT62 1401 741399 N/A N/A 85745 85761 AAGCATCTCCTGTGGTG 78 1402 741400 N/AN/A 85970 85986 CAACATGTTTTATCATG 68 1403 741401 N/A N/A 86195 86211AATATGACCACAATTTT 73 1404 741402 N/A N/A 86420 86436 TGGCAATATGAATGTGC86 1405 741403 N/A N/A 86645 86661 GATAAGGGCACATTGTC 64 1406 741404 N/AN/A 86871 86887 GTGATGGAGGGAAATCG 53 1407 741405 N/A N/A 87096 87112TGTGGTAATTGGAACAA 34 1408 741406 N/A N/A 87321 87337 ACTGAACCCAAATGGCT68 1409 741407 N/A N/A 87546 87562 TCACATTCATCATATTC 81 1410 741408 N/AN/A 87772 87788 CATTGCTGTTGTTGTTC 92 1411 741409 N/A N/A 87945 87961TAAGTTGTGACCATGCA 87 1412 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC95  402 741411 N/A N/A 87947 87963 AGTAAGTTGTGACCATG 96 1413 741412 N/AN/A 87949 87965 TTAGTAAGTTGTGACCA 76 1414 741413 N/A N/A 87997 88013AATAAAAATGTGCATGC 47 1415 741414 N/A N/A 88222 88238 AACAAAATACAGTCAGA91 1416 741415 N/A N/A 88447 88463 TTTGTACTGTGTGCTGT 89 1417 741416 N/AN/A 89657 89673 TGACATACAAACCCAAA 78 1418 N/A N/A 88652 88668 741417 N/AN/A 89658 89674 TTGACATACAAACCCAA 87  326 N/A N/A 88653 88669 741418 N/AN/A 89659 89675 CTTGACATACAAACCCA 82 1419 N/A N/A 88654 88670 741419 N/AN/A 89681 89697 GTCCCCAATCCCCACCC 19  100 N/A N/A 88676 88692 741420 N/AN/A 88705 88721 GAAGTTAACTCCCTAGA 83 1420 741421 N/A N/A 88707 88723ATGAAGTTAACTCCCTA 92 1421 741422 N/A N/A 89713 89729 GATGAAGTTAACTCCCT92  176 N/A N/A 88708 88724 741423 N/A N/A 89714 89730 AGATGAAGTTAACTCCC86 1422 N/A N/A 88709 88725 741424 N/A N/A 89716 89732 AGAGATGAAGTTAACTC78  252 N/A N/A 88711 88727 741425 N/A N/A 88754 88770 TTTTCAAGAGCTTTTCG67 1423 741426 N/A N/A 89761 89777 CCTTTTCAAGAGCTTTT 90 1424 N/A N/A88756 88772 741427 N/A N/A 89763 89779 TCCCTTTTCAAGAGCTT 96 1425 N/A N/A88758 88774 741428 N/A N/A 89765 89781 TTTCCCTTTTCAAGAGC 93 1426 N/A N/A88760 88776 741429 N/A N/A 89767 89783 TATTTCCCTTTTCAAGA 32 1427 N/A N/A88762 88778 741430 N/A N/A 88901 88917 ACAAGTAGGTAGGTCAA 83 1428 741431N/A N/A 89126 89142 TGAGCCATATTCAATAT 66 1429 741432 N/A N/A 89351 89367AAATTGCTAGGTTCAAC 77 1430 741433 N/A N/A 89579 89595 ATCAAATATTTACTAGA49 1431 741434 N/A N/A 89655 89671 ACATACAAACCCAAAGA 43 1432 N/A N/A88650 88666 741435 N/A N/A 89661 89677 CACTTGACATACAAACC 58 1433 N/A N/A88656 88672 741436 N/A N/A 89710 89726 GAAGTTAACTCCCTTGA 69 1434 741437N/A N/A 89712 89728 ATGAAGTTAACTCCCTT 92 1435 741438 N/A N/A 89759 89775TTTTCAAGAGCTTTTCT 66 1436 741439 N/A N/A 89804 89820 TCTACAGGTTATATGTG46 1437 741440 N/A N/A 90029 90045 TCCCAAAGTGCAAGACT 53 1438 741441 N/AN/A 90321 90337 CTCTATTGTTATATTTT 86 1439 741442 N/A N/A 90546 90562ATCTAACTCCTAGCACA 37 1440 741443 N/A N/A 90771 90787 ATACTTTCTCTGCATAA65 1441 741444 N/A N/A 91050 91066 TAGCTATAGTGCAATGG 52 1442 741445 N/AN/A 91277 91293 CTGGAATTCCAGAAAAA 63 1443 741446 N/A N/A 91502 91518CTTTCAAATCTCATTAC 68 1444 741447 N/A N/A 91727 91743 TCTTCTTTTGCAGAGAT65 1445 741448 N/A N/A 91952 91968 TAGAGCATTAAGAACAT 68 1446 741449 N/AN/A 92177 92193 GTTACTAAAAAAAACCA 41 1447 741450 N/A N/A 92402 92418TCCCATTGGACTGAGTT 53 1448 741451 N/A N/A 92627 92643 TATCCATTTTCCAGTTA83 1449 741452 N/A N/A 92852 92868 CCAGGGTGCTATACAAA 73 1450 741453 N/AN/A 93077 93093 CCTTAACAATCTTATTT 48 1451 741454 N/A N/A 93302 93318CACCACATTAATTAAAC 52 1452 741455 N/A N/A 93527 93543 ATGTTTTGAGTTCCAGG97 1453 741456 N/A N/A 93752 93768 TAATTAATAATCATCTT 20 1454 741457 N/AN/A 93950 93966 TTCTGGCTGACTGAATT 48 1455 741458 N/A N/A 93953 93969GGCTTCTGGCTGACTGA 72  180 741459 N/A N/A 93954 93970 TGGCTTCTGGCTGACTG83 1456 741460 N/A N/A 93956 93972 TGTGGCTTCTGGCTGAC 66 1457 741461 N/AN/A 93983 93999 GGCTTTTAACAAAACAA 69 1458 741462 N/A N/A 94052 94068ATAATTCAAGTCAGGGA 67 1459 741463 N/A N/A 94054 94070 CCATAATTCAAGTCAGG80 1460 741464 N/A N/A 94055 94071 GCCATAATTCAAGTCAG 89  331 741465 N/AN/A 94056 94072 TGCCATAATTCAAGTCA 66 1461 741466 N/A N/A 94058 94074ACTGCCATAATTCAAGT 64 1462 741467 N/A N/A 94208 94224 TAATATTGTGACCACTT94 1463 741468 N/A N/A 94433 94449 TAAGACTATTGCTTTGG 74 1464 741469 N/AN/A 94658 94674 CATAATAGATGAGTTAA 62 1465 741470 N/A N/A 94993 95009TTCAGTTTTGTGGCGGG 72 1466 741471 N/A N/A 95218 95234 ATTACATTAAAAGGTGG39 1467

TABLE 22 Percent reduction of human SNCA mRNA with 4-9-4 MOEand cEt gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 Sequence % ID No start stopstart stop (5′ to 3′) Reduction NO 740410 240 256   4691   4707GAATTCCTTTACACCAC 97   33 740432 270 286   4721   4737 AGTCCTTTCATGAATAC95  591 741472 N/A N/A  95443  95459 CCACCGTCACTGCATAC 85 1468 741473N/A N/A  95668  95684 ACTCTGTTGAATTTTCT 80 1469 741474 N/A N/A  95893 95909 TTTCCAGTGCTAGTATT 68 1470 741475 N/A N/A  96118  96134AATGAGATGAAAATTGA 70 1471 741476 N/A N/A  96343  96359 AGCTAGTTTGTAAACAA72 1472 741477 N/A N/A  96568  96584 AGAAGCAGTGAATCCAA 84 1473 741478N/A N/A  96793  96809 CTGTTAATCACCCCTTT 60 1474 741479 N/A N/A  97018 97034 CACAATACAGAGCAGAG 72 1475 741480 N/A N/A  97243  97259AGAAGTCAGACTTCAGG 33 1476 741481 N/A N/A  97474  97490 ATGGAAGATGAAAAAGG 3 1477 741482 N/A N/A  97699  97715 GTTGAGTCTGAGATGCC 75 1478 741483N/A N/A  97924  97940 AAGGCTGTTCACTATAT 81 1479 741484 N/A N/A  98149 98165 TCTTGCACTGATTCCTC 61 1480 741485 N/A N/A  98374  98390AGATAAGAAGCAAATGC 61 1481 741486 N/A N/A  98805  98821 GAATGGGCGGATCACAA34 1482 741487 N/A N/A  99032  99048 GATTATTTTAAGCACTT 90 1483 741488N/A N/A  99257  99273 AGAAAAAGGGCATTTAA 26 1484 741489 N/A N/A  99483 99499 TGCAATGTGTAGGTGGG 70 1485 741490 N/A N/A  99708  99724ACTTTTAAGGCATCCAT 74 1486 741491 N/A N/A  99933  99949 CCCTCCCAACAATTTCA26 1487 741492 N/A N/A 100158 100174 CTTTCCATTATTGTTCT 67 1488 741493N/A N/A 100391 100407 GGAAATGTTTATATATA 58 1489 741494 N/A N/A 100625100641 TAGGAAGTCTGGCTCCA 22 1490 741495 N/A N/A 100850 100866GATAATGGGCTAGGTGT 69 1491 741496 N/A N/A 101075 101091 TGGAATATCTTTGCTTA22 1492 741497 N/A N/A 101300 101316 ATAGCTTCAAGATCGGT 72 1493 741498N/A N/A 101525 101541 GAGATAAAGAGTCTGCT 61 1494 741499 N/A N/A 101803101819 TCACGGGATCACGCCAT 58 1495 741500 N/A N/A 102028 102044TGACTGAATAAGACATT 52 1496 741501 N/A N/A 102253 102269 GCAACAACTGCCAGCTT54 1497 741502 N/A N/A 102478 102494 CAGGTTTAAATACATTC 85 1498 741503N/A N/A 102703 102719 TTGGATAATCTGTTACT 63 1499 741504 N/A N/A 102968102984 TAATGCAGTGATACAAT 57 1500 741505 N/A N/A 103193 103209CTGGATCACTTGGGAAT 69 1501 741506 N/A N/A 103418 103434 TGTTCTAATTAAAAAGT47 1502 741507 N/A N/A 103643 103659 ACTTTACAACAAGATAA 36 1503 741508N/A N/A 103868 103884 TGATACATTATAATACA 58 1504 741509 N/A N/A 104093104109 GGGAAAGTATAGTTATG 63 1505 741510 N/A N/A 104332 104348GCATAAGAAAGAACAAT 42 1506 741511 N/A N/A 104557 104573 TCTTGAGGTCATAAATC56 1507 741512 N/A N/A 104782 104798 AAATGAAGGCGATAGAC 76 1508 741513N/A N/A 105007 105023 CTAAAAAAGAACTTTGA  0 1509 741514 N/A N/A 105232105248 TGTGTGATCAACTTTCA 88 1510 741515 N/A N/A 105457 105473AGTAAGCTTCAATTGGT 71 1511 741516 N/A N/A 105682 105698 AGGTTTCATCAATTATC89 1512 741517 N/A N/A 105907 105923 AGTGTCTTGTTAAGTAT 64 1513 741518N/A N/A 106134 106150 GAATTTACATAATCTTT 69 1514 741519 N/A N/A 106361106377 CTTTTTAAATAAACCTG 58 1515 741520 N/A N/A 106586 106602GAATAGCTGTAGACTTT 67 1516 741521 N/A N/A 106811 106827 TACCAATATAACAAATG23 1517 741522 N/A N/A 107037 107053 TATTTACTGTTTCATAA 40 1518 741523N/A N/A 107275 107291 TCAGGTGTCCTAGTGGG 68 1519 741524 N/A N/A 107500107516 GCAACCCCAAAATACTA 62 1520 741525 N/A N/A 107725 107741GTGTGATGATATATTGC 85 1521 741526 N/A N/A 107954 107970 ACAAGACAAAGAATACG47 1522 741527 N/A N/A 108273 108289 GTTCTCCTATAGTCCCA 24 1523 741528N/A N/A 108498 108514 ACTAGGGATGACAGCAC 74 1524 741529 N/A N/A 108724108740 TTCTTGCTTATATCAAT 72 1525 741530 N/A N/A 108970 108986GCAGTAATGGAACAGCG 63 1526 741531 N/A N/A 109195 109211 ATTTTGATATGGACCAG73 1527 741532 N/A N/A 109420 109436 TGCTGAGAAGTTTCCTA 52 1528 741533N/A N/A 109645 109661 TGCCCTTTTTATAAACT 18 1529 741534 N/A N/A 109870109886 AGCCTAAAGGGACTTGG 49 1530 741535 N/A N/A 110095 110111AGACTGAGACTATACAT 65 1531 741536 N/A N/A 110320 110336 GTCTATATTATAGATAC16 1532 741537 N/A N/A 110626 110642 TTACAATGAAACCCCAT 38 1533 741538N/A N/A 110853 110869 TTTACTATTTAGGAAAT 10 1534 741539 N/A N/A 111078111094 AAGTAAGAAGCACAAAA 20 1535 741540 N/A N/A 111303 111319AACTTGCAAGTTGTCCA 79 1536 741541 N/A N/A 111528 111544 GATTTCCCTAACTTTCC61 1537 741542 N/A N/A 111986 112002 ATGTCTCCTCTTCTGTT 43 1538 741543N/A N/A 112211 112227 GTAACCTGGCCACTTTG 49 1539 741544 N/A N/A 112436112452 TGTCTGTGTGAGACAGT 31 1540 741545 N/A N/A 112661 112677ATCATAATGAAGAAATG 12 1541 741546 N/A N/A 112886 112902 TGCCTTTGCTTCTGATA63 1542 741547 N/A N/A 113111 113127 ATATCAGGATTCTGCTT 52 1543 741548N/A N/A 113336 113352 GATGCTCTAATTCTCAG 61 1544

TABLE 23 Percent reduction of human SNCA mRNA with 4-9-4 MOEand cEt gapmers with mixed internucleoside linkages SEQ SEQ SEQ SEQ SEQSEQ ID ID ID ID ID ID SEQ Compound No: 3 No: 3 No: 5 No: 5 No: 6 No: 6Sequence % ID No start stop start stop start stop (5′ to 3′) ReductionNO 740914 N/A N/A 31  47 N/a N/a CACTCTTTCAGAGCTGG 49 1545 740915 N/AN/A 34  50 N/a N/a CCACACTCTTTCAGAGC 12 1546 740916 N/A N/A 37  53 N/aN/a ACACCACACTCTTTCAG 37 1547 740917 N/A N/A 40  56 N/a N/aTTTACACCACACTCTTT 55 1548 740918 N/A N/A 41  57 N/a N/aCTTTACACCACACTCTT 70 1549 740919 N/A N/A 43  59 98 114 TCCTTTACACCACACTC89 1550 740920 N/A N/A 90 106 90 106 ACCACACTCACTTCCGC 23 1551 740921N/A N/A 93 109 93 109 TACACCACACTCACTTC  0 1552 740922 N/A N/A 96 112 96112 CTTTACACCACACTCAC 86 1553 740923 370 386 N/A N/A N/A N/ACACTACATAGAGAACAC 86 1554 740924 373 389 N/A N/A N/A N/AAGCCACTACATAGAGAA  8 1555 740925 376 392 N/A N/A N/A N/ACTCAGCCACTACATAGA 29 1556 740926 379 395 N/A N/A N/A N/ACTTCTCAGCCACTACAT 52 1557 740927 382 398 N/A N/A N/A N/AGGTCTTCTCAGCCACTA 83 1558 740928 513 529 N/A N/A N/A N/ATCCTTGCCCAACTGGTC 46 1559 740929 516 532 N/A N/A N/A N/ACCTTCCTTGCCCAACTG 19 1560 740930 519 535 N/A N/A N/A N/ATACCCTTCCTTGCCCAA 50 1561 740931 522 538 N/A N/A N/A N/ATGATACCCTTCCTTGCC 16 1562 740932 525 541 N/A N/A N/A N/ATCTTGATACCCTTCCTT 63 1563

Example 4: Effect of 5-8-4 MOE and cEt Gapmers with MixedInternucleoside Linkages on Human SNCA In Vitro, Single Dose

Modified oligonucleotides complementary to a human SNCA nucleic acidwere designed and tested for their effect on SNCA mRNA in vitro. Themodified oligonucleotides were tested in a series of experiments thathad similar culture conditions.

Cultured SH-SY5Y cells at a density of 20,000 cells per well weretransfected using electroporation with 1,000 nM concentration ofmodified oligonucleotide or no modified oligonucleotide for untreatedcontrols. After approximately 24 hours, RNA was isolated from the cellsand SNCA mRNA levels were measured by quantitative real-time PCR usinghuman primer probe set RTS2621 as described in Example 1. SNCA mRNAlevels were adjusted according to total RNA content, as measured byRIBOGREEN®. Results are presented in the tables below as percentreduction of the amount of SNCA mRNA, relative to untreated controlcells.

The modified oligonucleotides in tables 24-28 are 4-9-4 MOE and cEtgapmers. The gapmers are 17 nucleobases in length, wherein the centralgap segment comprises nine 2′-deoxynucleosides and is flanked by wingsegments on both the 5′ end on the 3′ end comprising two 2′-MOEnucleosides and two cEt nucleosides. The sugar motif for the gapmers is(from 5′ to 3′): eekkdddddddddkkee; wherein ‘d’ represents a2′-deoxyribose sugar; ‘e’ represents a 2′-MOE modified sugar; and ‘k’represents a cEt modified sugar. All cytosine residues throughout eachgapmer are 5-methyl cytosines. The internucleoside linkages are mixedphosphodiester and phosphorothioate linkages. The internucleosidelinkage motif for the gapmers is (from 5′ to 3′): sooosssssssssoss;wherein ‘o’ represents a phosphodiester internucleoside linkage and ‘s’represents a phosphorothioate internucleoside linkage. “Start Site”indicates the 5′-most nucleoside to which the gapmer is complementary inthe human nucleic acid sequence. “Stop Site” indicates the 3′-mostnucleoside to which the gapmer is complementary in the human nucleicacid sequence.

Each modified oligonucleotide listed in the Tables below iscomplementary to human SNCA nucleic acid sequences SEQ ID NO: 1, SEQ IDNO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6, asindicated. ‘N/A’ indicates that the modified oligonucleotide is notcomplementary to that particular nucleic acid with 100% complementarity.A value of 0% reduction indicates that the compound had no effect orincreased mRNA concentrations in the cell. As shown below, modifiedoligonucleotides complementary to human SNCA reduced the amount of humanSNCA mRNA.

TABLE 24 Percent reduction of human SNCA mRNA with 4-9-4 MOEand cEt gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 Sequence % ID No start stopstart stop (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 61   33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC87  591 741010 N/A N/A 20549 20565 CAATTTCTAGGTTCTAT 28 1107 741011 N/AN/A 20559 20575 AACTATGCTGCAATTTC 28 1108 741012 N/A N/A 20561 20577ACAACTATGCTGCAATT 25 1109 741013 N/A N/A 20562 20578 CACAACTATGCTGCAAT54  314 741014 N/A N/A 20565 20581 TTCCACAACTATGCTGC 51 1110 741015 N/AN/A 20774 20790 GACCACAATTGCAGACA 55 1111 741016 N/A N/A 20985 21001GTGTGAGCAAACATTCT 73 1112 741017 N/A N/A 27412 27428 CAGTGTGAGCAAACATT35 1113 N/A N/A 20987 21003 741018 N/A N/A 27413 27429 ACAGTGTGAGCAAACAT67  468 N/A N/A 20988 21004 741019 N/A N/A 27414 27430 CACAGTGTGAGCAAACA61 1114 N/A N/A 20989 21005 741020 N/A N/A 20991 21007 GGCACAGTGTGAGCAAA53 1115 741021 N/A N/A 20999 21015 AAGTTTCTGGCACAGTG 89 1116 741022 N/AN/A 21224 21240 GTTCAGAATTATGTCAT 81 1117 741023 N/A N/A 21449 21465TCTTATGTGCACATGAG 16 1118 741024 N/A N/A 21674 21690 CATAGTAGCATTACAGA47 1119 741025 N/A N/A 21899 21915 TCAGGCAGTGGCTTCAC 43 1120 741026 N/AN/A 22129 22145 TAAAAAAAGTTGTTCAT  0 1121 741027 N/A N/A 22360 22376CACTCAAGTGTTTAAAA 26 1122 741028 N/A N/A 22454 22470 TGTGACCTGTGCTTGTT83 1123 741029 N/A N/A 22456 22472 CCTGTGACCTGTGCTTG 87 1124 741030 N/AN/A 22457 22473 GCCTGTGACCTGTGCTT 62   88 741031 N/A N/A 22458 22474TGCCTGTGACCTGTGCT 54 1125 741032 N/A N/A 22460 22476 GTTGCCTGTGACCTGTG78 1126 741033 N/A N/A 22599 22615 TATTAGACACTTAAGGG 28 1127 741034 N/AN/A 22831 22847 TCAATCTTAAATTTTTC 56 1128 741035 N/A N/A 23056 23072GTACTTTCCCACCTAGA 39 1129 741036 N/A N/A 23281 23297 TCTCAGAGACCACAGCT56 1130 741037 N/A N/A 23285 23301 TTGTTCTCAGAGACCAC 86 1131 741038 N/AN/A 23286 23302 ATTGTTCTCAGAGACCA 72  164 741039 N/A N/A 23287 23303TATTGTTCTCAGAGACC 67 1132 741040 N/A N/A 23289 23305 CATATTGTTCTCAGAGA40 1133 741041 N/A N/A 23506 23522 ACTATTAACCACTGATC 25 1134 741042 N/AN/A 23731 23747 GTTGCAGTCCACAGAAT 34 1135 741043 N/A N/A 23956 23972TAAAGATAAGTATCTCA 70 1136 741044 N/A N/A 24181 24197 AAAACAAACCTAAGTCA 0 1137 741045 N/A N/A 24406 24422 AAAAGCTAACAGCCTAT 14 1138 741046 N/AN/A 24631 24647 TTAAATTGATGAGATGT 49 1139 741047 N/A N/A 24856 24872GTATTCTTTGCATTAGT 67 1140 741048 N/A N/A 25081 25097 TAAAAGTGTACATTATT22 1141 741049 N/A N/A 25306 25322 CTCAAGGCAAAGCTGTA 57 1142 741050 N/AN/A 25531 25547 TGCCACTATAAGCAGTC 55 1143 741051 N/A N/A 25756 25772TTCAAGCCCATGCCCTC 21 1144 741052 N/A N/A 25801 25817 ATCCAGTAGAGTGAGAG37 1145 741053 N/A N/A 25803 25819 TCATCCAGTAGAGTGAG 41 1146 741054 N/AN/A 25804 25820 ATCATCCAGTAGAGTGA 31  315 741055 N/A N/A 25807 25823GACATCATCCAGTAGAG 55 1147 741056 N/A N/A 25923 25939 TGAATACATTGTCTTAA18 1148 741057 N/A N/A 25925 25941 ATTGAATACATTGTCTT 41 1149 741058 N/AN/A 25926 25942 AATTGAATACATTGTCT 50  392 741059 N/A N/A 25927 25943TAATTGAATACATTGTC 29 1150 741060 N/A N/A 25929 25945 CATAATTGAATACATTG25 1151 741061 N/A N/A 25981 25997 TGAGTAGCTATGGTTTA 37 1152 741062 N/AN/A 26202 26218 TCTTTGTGTTATACAAT  0 1153 741063 N/A N/A 26204 26220CCTCTTTGTGTTATACA 53 1154 741064 N/A N/A 26205 26221 CCCTCTTTGTGTTATAC42  469 741065 N/A N/A 26206 26222 TCCCTCTTTGTGTTATA 20 1155 741066 N/AN/A 26208 26224 TTTCCCTCTTTGTGTTA 30 1156 741067 N/A N/A 26431 26447TACATACAATATTAAGG  0 1157 741068 N/A N/A 26656 26672 AAAAGAATGGATTCTGA34 1158 741069 N/A N/A 26881 26897 AAGGAAAAACTCTGCCC 15 1159 741070 N/AN/A 27106 27122 TCACCCCAAGGCATTTG  6 1160 741071 N/A N/A 27331 27347ACACCCTGATTCCCAAG 31 1161 741072 N/A N/A 27410 27426 GTGTGAGCAAACATTCA52 1162 741073 N/A N/A 27416 27432 GTCACAGTGTGAGCAAA 72 1163 741074 N/AN/A 27556 27572 GGGAAGTATTAGTGGAA 27 1164 741075 N/A N/A 27782 27798GCTGAAAATATGAAACA 32 1165 741076 N/A N/A 28007 28023 ACTTCTAGCACTATTTT 9 1166 741077 N/A N/A 28232 28248 TTGTGCATTTATTCCAC 78 1167 741078 N/AN/A 28457 28473 GACTGTAATCTAGGACC 74 1168 741079 N/A N/A 28682 28698TGACTTTTGAATCAGTC 14 1169 741080 N/A N/A 29010 29026 GAGCGATTCTCCTGGTT61 1170 741081 N/A N/A 29235 29251 CACAGTCCATAATATTG 34 1171 741082 N/AN/A 29460 29476 TTTTTGTTAATAGTTCT 73 1172 741083 N/A N/A 29685 29701GCTTTCTCAGAGCCCAA 74 1173 741084 N/A N/A 29912 29928 ATCTCTCTACCATGTGA34 1174 741085 N/A N/A 30137 30153 GTGGATAAAGTACATTA 16 1175 741086 N/AN/A 30362 30378 AAATGGTATTCAGAGAT 42 1176

TABLE 25 Percent reduction of human SNCA mRNA with 4-9-4 MOEand cEt gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 Sequence % ID No start stopstart stop (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 55   33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC84  591 741087 N/A N/A 30587 30603 TTTTCTCCTAAAGCCTT 42 1177 741088 N/AN/A 31037 31053 CAGATTTCCAGCACACT 30 1178 741089 N/A N/A 31262 31278CCTTCTTAGTGGTAAGA  0 1179 741090 N/A N/A 31487 31503 AATTACAGTGTAGGTAA18 1180 741091 N/A N/A 31712 31728 ATAAGAGGTCACTGGAT 25 1181 741092 N/AN/A 31937 31953 AAGGAAACAGTCTACAT 14 1182 741093 N/A N/A 32162 32178CTATCATGATAAGTATA 10 1183 741094 N/A N/A 32387 32403 TGTGGTTCTGCCCATCT54 1184 741095 N/A N/A 32624 32640 GCCTAAACATTTTACTT  5 1185 741096 N/AN/A 32858 32874 GAAGTTTCTGAAGAAAT 40 1186 741097 N/A N/A 33083 33099TTTTCAGTAGATTTGAC 24 1187 741098 N/A N/A 33308 33324 GCTATGACCCTCAAGCC11 1188 741099 N/A N/A 33533 33549 AATAGAGCAAAATTTCG 35 1189 741100 N/AN/A 33762 33778 ATAATCAAACAAAAGGG 16 1190 741101 N/A N/A 33987 34003AAAGTTCAATGCTGTGT 69 1191 741102 N/A N/A 34212 34228 GAAATGGGCATGTAAAC10 1192 741103 N/A N/A 34443 34459 CAAAATACAATGTTCAA 15 1193 741104 N/AN/A 34668 34684 ATTCTTCTATCCTAGAA  5 1194 741105 N/A N/A 34893 34909ATTATCATGGTTGCCCA 38 1195 741106 N/A N/A 35118 35134 ATGAGATCTTTTTGCAT39 1196 741107 N/A N/A 35343 35359 AAGCAAGTTGTCCATGG 47 1197 741108 N/AN/A 35568 35584 TGTTGGAGTTTACAATT 20 1198 741109 N/A N/A 35793 35809CTCACTAGCCCTGTGAC  0 1199 741110 N/A N/A 36018 36034 TCTCTTTCATGGGTATT57 1200 741111 N/A N/A 36252 36268 GTCATTTTAATAAGTGT 65 1201 741112 N/AN/A 36484 36500 CAATTAAATAAACCTCT 10 1202 741113 N/A N/A 36790 36806TATGGTGATATGGTTAG 53 1203 741114 N/A N/A 37018 37034 CCATGTGTTTTTGTGGC33 1204 741115 N/A N/A 37243 37259 CAAAGGTATAAGGTCAT 49 1205 741116 N/AN/A 37468 37484 AGCTTGTATTTTTGAAA 24 1206 741117 N/A N/A 37788 37804CGCATCTGTCTTTCTTT 25 1207 741118 N/A N/A 38013 38029 TAGGACAGGTGAAATAA12 1208 741119 N/A N/A 38238 38254 AGTTATTAGAATAACAC  0 1209 741120 N/AN/A 38464 38480 AATAAAATGTCTTAATC  0 1210 741121 N/A N/A 38691 38707ACTCAAAAAAGAAGAAT  0 1211 741122 N/A N/A 38916 38932 GTTTTCTCTGTATTGGC87 1212 741123 N/A N/A 39141 39157 TGGCCTAGTGGTTATAA  0 1213 741124 N/AN/A 39366 39382 CACAAAGAGGAAACAGG 27 1214 741125 N/A N/A 39591 39607ACATTTTTTAACTGGAT 76 1215 741126 N/A N/A 39816 39832 AGGCTAAATTTTAATAA 0 1216 741127 N/A N/A 40041 40057 TAGCCTTTCATAGTACG 38 1217 741128 N/AN/A 40266 40282 AAGAGGAAAAGCTTGGA  0 1218 741129 N/A N/A 40491 40507AAAAATTCTGGTGCCAA 57 1219 741130 N/A N/A 40716 40732 AAGCTAAACTACCGCTG 2 1220 741131 N/A N/A 40941 40957 GAATTTCCTGGATGCTC 44 1221 741132 N/AN/A 41130 41146 AGATTCCAGCAGAGATT 20 1222 741133 N/A N/A 41132 41148ACAGATTCCAGCAGAGA 44 1223 741134 N/A N/A 41133 41149 AACAGATTCCAGCAGAG40   90 741135 N/A N/A 41134 41150 GAACAGATTCCAGCAGA 24 1224 741136 N/AN/A 41136 41152 GTGAACAGATTCCAGCA 34 1225 741137 N/A N/A 41166 41182ATCTGTAAGAAGTTTAG 10 1226 741138 N/A N/A 41391 41407 TGAGAAATTTTATGGGT47 1227 741139 N/A N/A 41620 41636 TCATTCAAAACCATCCT 21 1228 741140 N/AN/A 41845 41861 GATCACACTGCTTATAG 16 1229 741141 N/A N/A 42070 42086CAAGTTGATGGCATATA 34 1230 741142 N/A N/A 42295 42311 GTGTACCAACCTCAAGT34 1231 741143 N/A N/A 42532 42548 TAAGTAAATACCTAGGG 20 1232 741144 N/AN/A 42757 42773 GATTTGTGCCTGGCATC 38 1233 741145 N/A N/A 42835 42851TGCCTCTACCTCCAGCA 39 1234 741146 N/A N/A 42837 42853 GATGCCTCTACCTCCAG42 1235 741147 N/A N/A 42838 42854 TGATGCCTCTACCTCCA 40  166 741148 N/AN/A 42839 42855 CTGATGCCTCTACCTCC 33 1236 741149 N/A N/A 42982 42998TATCACAACTACATTGT  0 1237 741150 N/A N/A 43208 43224 GGCCTCCTGCTGCAGCA 0 1238 741151 N/A N/A 43440 43456 GCACTCATTTTAAATGT 20 1239 741152 N/AN/A 43665 43681 TGGTAACTTAGGACAAG 44 1240 741153 N/A N/A 43818 43834TTCTCTGGACCTCTTAA  6 1241 741154 N/A N/A 43820 43836 ACTTCTCTGGACCTCTT41 1242 741155 N/A N/A 43821 43837 TACTTCTCTGGACCTCT 49  242 741156 N/AN/A 43822 43838 TTACTTCTCTGGACCTC 44 1243 741157 N/A N/A 43890 43906TCAATACAACTTAATTC  0 1244 741158 N/A N/A 44376 44392 TTGGGCTGGAAGCAGTG 9 1245 741159 N/A N/A 44601 44617 AAGATATGCAGAGGGTT 49 1246 741160 N/AN/A 44828 44844 TGGTCTAACTGTGTTGC 40 1247 741161 N/A N/A 45053 45069GTTTATGGACTTTTTAA 29 1248 741162 N/A N/A 45278 45294 TTTTGTACTTTATGGAA40 1249 741163 N/A N/A 45503 45519 ACTTCTCCTTCAATTAA 11 1250

TABLE 26 Percent reduction of human SNCA mRNA with 4-9-4 MOEand cEt gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 Sequence % ID No start stopstart stop (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 64   33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC82  591 741164 N/A N/A 45728 45744 GTCAAAATATTCTTACT 30 1564 741165 N/AN/A 45953 45969 CACATAAAATTAAAGCT  2 1565 741166 N/A N/A 46157 46173TCCATGAAGCCAAGTAA 38 1566 741167 N/A N/A 46159 46175 TTTCCATGAAGCCAAGT74 1567 741168 N/A N/A 53645 53661 ATTTCCATGAAGCCAAG 67  317 N/A N/A46160 46176 741169 N/A N/A 46161 46177 GATTTCCATGAAGCCAA 84 1568 741170N/A N/A 46163 46179 GAGATTTCCATGAAGCC 87 1569 741171 N/A N/A 46178 46194GGAATTGGAGTGAGAGA 29 1570 741172 N/A N/A 46403 46419 ATCCCTACATACTCACA20 1571 741173 N/A N/A 46628 46644 TTCTACCACCCACAGCT  0 1572 741174 N/AN/A 46880 46896 GAAAACATTGTATTATT 39 1573 741175 N/A N/A 47105 47121CCTTAAAATGATGCCTG 43 1574 741176 N/A N/A 47330 47346 CTAAAGTTAAGGTGTCG25 1575 741177 N/A N/A 47557 47573 GCATGAATTACTTTACG 40 1576 741178 N/AN/A 47952 47968 GGTTGTTCAAGTGATTC 58 1577 741179 N/A N/A 48177 48193GATCTTTTCATCATGCC 75 1578 741180 N/A N/A 48225 48241 GGTCATGACTCTGACAC14 1579 741181 N/A N/A 48227 48243 CTGGTCATGACTCTGAC 34 1580 741182 N/AN/A 48228 48244 CCTGGTCATGACTCTGA 39  394 741183 N/A N/A 48229 48245CCCTGGTCATGACTCTG 46 1581 741184 N/A N/A 48231 48247 TCCCCTGGTCATGACTC35 1582 741185 N/A N/A 48402 48418 AGGGCCATCCTGTTCAA  9 1583 741186 N/AN/A 48648 48664 AGAATACTTATTTTTTG 14 1584 741187 N/A N/A 48713 48729ATTTTGGATGCTTCTGA 56 1585 741188 N/A N/A 48715 48731 GTATTTTGGATGCTTCT70 1586 741189 N/A N/A 48716 48732 TGTATTTTGGATGCTTC 80  471 741190 N/AN/A 48717 48733 TTGTATTTTGGATGCTT 78 1587 741191 N/A N/A 48719 48735GTTTGTATTTTGGATGC 69 1588 741192 N/A N/A 48873 48889 TTTAAAGATGGATATTG 0 1589 741193 N/A N/A 49111 49127 TAAGGTCCCTCCCTCAA  0 1590 741194 N/AN/A 49373 49389 TTACCTGGCTACCTTTT 31 1591 741195 N/A N/A 49480 49496TGAAATTTTCCAGCTAT 60 1592 741196 N/A N/A 80992 81008 TTGAAATTTTCCAGCTA37  167 N/A N/A 49481 49497 741197 N/A N/A 49482 49498 ATTGAAATTTTCCAGCT60 1593 741198 N/A N/A 49484 49500 TGATTGAAATTTTCCAG 33 1594 741199 N/AN/A 49598 49614 TGGGAATCACCTCCCCT 14 1595 741200 N/A N/A 49825 49841CATTGAATTAATTTGTT 30 1596 741201 N/A N/A 50050 50066 CACCATTTTATAGCATG64 1597 741202 N/A N/A 50275 50291 TGGAAAGAGGTATGAGT  0 1598 741203 N/AN/A 50500 50516 ATTAAAATGAGAGGTCC 18 1599 741204 N/A N/A 50725 50741TTCCACCACACAAGTTA 43 1600 741205 N/A N/A 50920 50936 TTCATCAATATCTGCAA66 1601 741206 N/A N/A 50921 50937 TTTCATCAATATCTGCA 85  243 741207 N/AN/A 50922 50938 TTTTCATCAATATCTGC 86 1602 741208 N/A N/A 50924 50940GGTTTTCATCAATATCT 76 1603 741209 N/A N/A 50950 50966 CTTTGATGAATTAAGAG22 1604 741210 N/A N/A 51175 51191 AGGTATAAGATTCCTGC 31 1605 741211 N/AN/A 51412 51428 ACAAGGCCTTACTTACG  9 1606 741212 N/A N/A 51637 51653CTGCCCAACTTACAATT  9 1607 741213 N/A N/A 51868 51884 CATGGCAAGAACAAGGG27 1608 741214 N/A N/A 52093 52109 TATTATGTGCTTATTGG 56 1609 741215 N/AN/A 52318 52334 CCTAACACATGGATGTA  5 1610 741216 N/A N/A 52417 52433CAAATGTATAGAGAAGT 15 1611 741217 N/A N/A 52419 52435 ATCAAATGTATAGAGAA41 1612 741218 N/A N/A 52420 52436 GATCAAATGTATAGAGA 36  395 741219 N/AN/A 52421 52437 AGATCAAATGTATAGAG 31 1613 741220 N/A N/A 52423 52439ACAGATCAAATGTATAG 55 1614 741221 N/A N/A 52543 52559 CTCTACTGTGTTTGAGC14 1615 741222 N/A N/A 52768 52784 CTATATACTACAATTTT  7 1616 741223 N/AN/A 52993 53009 TGAGCTCACTGACAGAA 13 1617 741224 N/A N/A 53239 53255CAAGTATAAATATGTTT 10 1618 741225 N/A N/A 53464 53480 CCAAGGAGCATTTGGAT 7 1619 741226 N/A N/A 53642 53658 TCCATGAAGCCAAGATC 52 1620 741227 N/AN/A 53644 53660 TTTCCATGAAGCCAAGA 65 1621 741228 N/A N/A 53646 53662TATTTCCATGAAGCCAA 79 1622 741229 N/A N/A 53648 53664 ATTATTTCCATGAAGCC81 1623 741230 N/A N/A 53689 53705 ATCCATAAATGCTTTGT 68 1624 741231 N/AN/A 53914 53930 TATCTTCATCATAGCTC 60 1625 741232 N/A N/A 54139 54155AGACCACACTCCAACTA 20 1626 741233 N/A N/A 54364 54380 ATGTAAGGATGATCATT32 1627 741234 N/A N/A 54589 54605 TGACTTTATATGCATTT 54 1628 741235 N/AN/A 54814 54830 CATATATACTTACTTAC  2 1629 741236 N/A N/A 55039 55055ATATGTTTGATCGAAAG 20 1630 741237 N/A N/A 55269 55285 CAGATGGTTTTTTCTTT31 1631 741238 N/A N/A 55494 55510 ACAAAAGGGATTGTTCT  8 1632 741239 N/AN/A 55719 55735 ACTTGACTATAACACTT 40 1633 741240 N/A N/A 56172 56188ATAGAAAACAGATGAAG  2 1634

TABLE 27 Percent reduction of human SNCA mRNA with 4-9-4 MOEand cEt gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 Sequence % ID No start stopstart stop (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 57   33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC87  591 741241 N/A N/A 56397 56413 AAAATTTTTGCACACTT 46 1251 741242 N/AN/A 56622 56638 GCCAAATCAATGGATGA 33 1252 741243 N/A N/A 56847 56863AGTGACCAAGAGAATGA  4 1253 741244 N/A N/A 57072 57088 TTTTAAAACACTGGCCT 0 1254 741245 N/A N/A 57297 57313 TAGGATTAAACAGTCCA  0 1255 741246 N/AN/A 57522 57538 TTATCTGTTGCTATGTG 51 1256 741247 N/A N/A 57747 57763AAGAAGGAGAATAGCAG 14 1257 741248 N/A N/A 57981 57997 CGGGCAAACATGTTTTG 8 1258 741249 N/A N/A 58206 58222 ATGACCTACATGCTAAA 11 1259 741250 N/AN/A 58431 58447 AGAAGCAAAATGTCAGT 40 1260 741251 N/A N/A 58656 58672CCTAACAGCTTTACTTT  0 1261 741252 N/A N/A 58881 58897 CTTTCACACATCTCTAA 0 1262 741253 N/A N/A 58991 59007 TTTCATTAATCTGTGAA 14 1263 741254 N/AN/A 58992 59008 ATTTCATTAATCTGTGA 20  169 741255 N/A N/A 58993 59009TATTTCATTAATCTGTG 39 1264 741256 N/A N/A 58995 59011 TATATTTCATTAATCTG27 1265 741257 N/A N/A 59106 59122 CCTTACACAAAATATAA  0 1266 741258 N/AN/A 59354 59370 ACACCAATATATTATTT 13 1267 741259 N/A N/A 59594 59610TAAAGGATGCAAAGGCA  0 1268 741260 N/A N/A 59948 59964 TTCCAGCGATCCCACTC21 1269 741261 N/A N/A 60173 60189 CTCAACATCTTTAATGA  6 1270 741262 N/AN/A 60421 60437 GGGACCTAAAACTATAA  0 1271 741263 N/A N/A 60758 60774AGCAGAATAGAAAATCC 14 1272 741264 N/A N/A 60983 60999 TTCAATGCGACTCCCAT23 1273 741265 N/A N/A 61216 61232 CAACAAAACTGAGAATC  0 1274 741266 N/AN/A 61474 61490 AATGCCTGCTTTCACCA 26 1275 741267 N/A N/A 61699 61715TATAAGCAGGAGTAAAA  2 1276 741268 N/A N/A 61969 61985 GTTCCAAAAGATAGAGA11 1277 741269 N/A N/A 62200 62216 CGTACACAAACTAGAAA  1 1278 741270 N/AN/A 62492 62508 TACTGTTGCATTCCAGC  6 1279 741271 N/A N/A 62729 62745TCTTAGTGTGGTGGCTC 15 1280 741272 N/A N/A 62955 62971 TCAACAATAATAATGAC 0 1281 741273 N/A N/A 63197 63213 CCTTTTCATCAACACAT 12 1282 741274 N/AN/A 63422 63438 TATGCATCTAACACTTG  8 1283 741275 N/A N/A 63666 63682CCATCAACCAAGTATCT  0 1284 741276 N/A N/A 63891 63907 CTTGAAACAGTAACTTG 0 1285 741277 N/A N/A 64116 64132 AACATAGCAGATTAATA 12 1286 741278 N/AN/A 64349 64365 TCATGTTATATAGTGGG 73 1287 741279 N/A N/A 64574 64590TGTAACCTAATGTAAAT  0 1288 741280 N/A N/A 64799 64815 ACAAGTATCTGTACTCA59 1289 741281 N/A N/A 65024 65040 GTCTCTGTTAATGTTGG 26 1290 741282 N/AN/A 65249 65265 GAACCAGCCTGACTTAA 21 1291 741283 N/A N/A 65474 65490TTGTATGGGTTACATAA  3 1292 741284 N/A N/A 65801 65817 CAATTAAATGCAATTCC 0 1293 741285 N/A N/A 66026 66042 TGACAGAAGTGTGCATA 14 1294 741286 N/AN/A 66251 66267 CAACACATCCACATTGC  8 1295 741287 N/A N/A 66476 66492TTCACACCTCTCTCCCT  0 1296 741288 N/A N/A 66701 66717 TGCTGGTCTAAGATGCA24 1297 741289 N/A N/A 66926 66942 ATGTGTTTTGAGGAAAA 13 1298 741290 N/AN/A 67151 67167 CAGAAGTAAATGTGGAC 24 1299 741291 N/A N/A 67376 67392TGATTCTTTGGATTCAT 27 1300 741292 N/A N/A 67876 67892 CATTCTTGTTTTTATTC37 1301 741293 N/A N/A 68101 68117 AATAGTGTCCCAGTGTA 40 1302 741294 N/AN/A 68326 68342 TGAAAGCTGTTCAGTTA 12 1303 741295 N/A N/A 68551 68567CCCACATATACTACTTG 32 1304 741296 N/A N/A 68776 68792 AGAATTTCAGGAAGTTA33 1305 741297 N/A N/A 68798 68814 CAAAGTAAGAGGAGATT 13 1306 741298 N/AN/A 68800 68816 GCCAAAGTAAGAGGAGA 37 1307 741299 N/A N/A 68801 68817TGCCAAAGTAAGAGGAG 11  397 741300 N/A N/A 68804 68820 CAGTGCCAAAGTAAGAG 0 1308 741301 N/A N/A 69001 69017 TGAATCCATTTGTCCAG 52 1309 741302 N/AN/A 69227 69243 CTCTAAAATACAAATGT 13 1310 741303 N/A N/A 69452 69468GAACAAAGGAATAAGTA  0 1311 741304 N/A N/A 69677 69693 CTAGATGTAGATATCAT13 1312 741305 N/A N/A 69902 69918 AAGGGAATAAATTGTAG 28 1313 741306 N/AN/A 70127 70143 CAACAGACCCTTTCAAT  3 1314 741307 N/A N/A 70352 70368GTCTTCCCACTGCCTAC  7 1315 741308 N/A N/A 70577 70593 TTTAGATATACCTCCAA37 1316 741309 N/A N/A 70880 70896 GCTTCAGTTTCTTGAGT 18 1317 741310 N/AN/A 71105 71121 CTGGTCTTTCTCACAAT  8 1318 741311 N/A N/A 71375 71391ATCATTCTTAACAGAAA 15 1319 741312 N/A N/A 71600 71616 GCTCTTGCTGTGCAGCC11 1320 741313 N/A N/A 71844 71860 ATTTAAAGCAGCAGTCC  4 1321 741314 N/AN/A 72076 72092 AGGTAATTCTAATTTTA 17 1322 741315 N/A N/A 72301 72317GGCAAATGACAGGGTCT 69 1323 741316 N/A N/A 72632 72648 TCTCAACTGCCTGAGTA 0 1324 741317 N/A N/A 72857 72873 CATGTCAGCTTTTTAGT 19 1325

TABLE 28 Percent reduction of human SNCA mRNA with 4-9-4 MOEand cEt gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 Sequence % ID No start stopstart stop (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 57   33 740432 270 286  4721  4737 AGTCCTTTCATGAATAC85  591 741318 N/A N/A 73090 73106 ATACTCAGATATTTAAA  0 1326 741319 N/AN/A 73188 73204 ACTTTCTGTGTGGTATG 42 1327 741320 N/A N/A 73190 73206AGACTTTCTGTGTGGTA 59 1328 741321 N/A N/A 73191 73207 CAGACTTTCTGTGTGGT83  170 741322 N/A N/A 73192 73208 ACAGACTTTCTGTGTGG 37 1329 741323 N/AN/A 73194 73210 AGACAGACTTTCTGTGT  0 1330 741324 N/A N/A 73315 73331GTTGAGAATTTTTCATT 14 1331 741325 N/A N/A 73540 73556 AGTTATGGAGCATCTTT46 1332 741326 N/A N/A 73765 73781 GACTGAGTTTTTTATTC 16 1333 741327 N/AN/A 73990 74006 TCCTGAATTAAAAATTT  0 1334 741328 N/A N/A 74215 74231GCTAAGCACAAACAATT 15 1335 741329 N/A N/A 74292 74308 GAACTCTGTAGTCAGAA58 1336 741330 N/A N/A 74294 74310 TAGAACTCTGTAGTCAG 63 1337 741331 N/AN/A 74295 74311 ATAGAACTCTGTAGTCA 42  398 741332 N/A N/A 74296 74312AATAGAACTCTGTAGTC 42 1338 741333 N/A N/A 74298 74314 TGAATAGAACTCTGTAG25 1339 741334 N/A N/A 74440 74456 ACACAGAGCACTTCTTA 15 1340 741335 N/AN/A 74665 74681 GGAGTTACAGAGTTGCC 64 1341 741336 N/A N/A 74890 74906TATCAGTCTATTAAGAA 11 1342 741337 N/A N/A 75115 75131 AAGTTTCTCAGAGCCTG24 1343 741338 N/A N/A 75340 75356 AATACAGAAGTCTATTC  0 1344 741339 N/AN/A 75573 75589 CATTGAATAAAAATTTG  0 1345 741340 N/A N/A 75945 75961CAGGTATAAAATTTTTT  2 1346 741341 N/A N/A 76170 76186 GGTGTTAATCACTTGAA18 1347 741342 N/A N/A 76398 76414 TCTTGAAGCTAGTTGGG 39 1348 741343 N/AN/A 76623 76639 AGGGCAACTAACCAACA 20 1349 741344 N/A N/A 76848 76864GTGGATACTTAGTATCA 13 1350 741345 N/A N/A 77073 77089 CTCTCTCAGTTGTAGGT19 1351 741346 N/A N/A 77298 77314 AAAGTATGCTGTGTTCT 46 1352 741347 N/AN/A 77523 77539 GTACCCGGCACTTTTCC 15 1353 741348 N/A N/A 77663 77679TCTAGAAAAGCTCTCTT  0 1354 741349 N/A N/A 77665 77681 ACTCTAGAAAAGCTCTC13 1355 741350 N/A N/A 77666 77682 GACTCTAGAAAAGCTCT 36  247 741351 N/AN/A 77667 77683 AGACTCTAGAAAAGCTC 26 1356 741352 N/A N/A 77748 77764TGGCACCCAGGAGTAAG  8 1357 741353 N/A N/A 77973 77989 CATACACAAAATCCCCT28 1358 741354 N/A N/A 78198 78214 CACATGAAGCCAGGGAC 19 1359 741355 N/AN/A 78423 78439 GCAGGCCCTAAACTGTG  5 1360 741356 N/A N/A 78648 78664AAATTTATCTATCATGC 30 1361 741357 N/A N/A 78873 78889 GCTAAACACTTTATCAA22 1362 741358 N/A N/A 79098 79114 ACTTCATTCTTTCTGTT 30 1363 741359 N/AN/A 79323 79339 CAATTAAAAGATTACTT  0 1364 741360 N/A N/A 79548 79564ACATTGTACAGTTAATT  9 1365 741361 N/A N/A 79773 79789 TACAAACCTTACTATGC 9 1366 741362 N/A N/A 79998 80014 AACAGACTTAAACAAAC 40 1367 741363 N/AN/A 80223 80239 CTCAGACATCATGTTTT 52 1368 741364 N/A N/A 80448 80464AGGCACTCACAAACATT 33 1369 741365 N/A N/A 80673 80689 TCTCGCATCCTAAATGT 0 1370 741366 N/A N/A 80898 80914 TTCATATTTTATGTTAC 23 1371 741367 N/AN/A 80991 81007 TGAAATTTTCCAGCTAA 52 1372 741368 N/A N/A 80993 81009CTTGAAATTTTCCAGCT 72 1373 741369 N/A N/A 80995 81011 ATCTTGAAATTTTCCAG33 1374 741370 N/A N/A 81123 81139 CTATAATTACATTCCTA  9 1375 741371 N/AN/A 81348 81364 GCATGAACCTAGATATG  4 1376 741372 N/A N/A 81472 81488GCTGTTTGAAGTGACAA 21 1377 741373 N/A N/A 81474 81490 GAGCTGTTTGAAGTGAC54 1378 741374 N/A N/A 81475 81491 AGAGCTGTTTGAAGTGA 52  249 741375 N/AN/A 81476 81492 GAGAGCTGTTTGAAGTG 22 1379 741376 N/A N/A 81478 81494TGGAGAGCTGTTTGAAG 12 1380 741377 N/A N/A 81575 81591 CTGCCACTATTCACAAT27 1381 741378 N/A N/A 81800 81816 TTATTGCATTAATGGAA 76 1382 741379 N/AN/A 82107 82123 ATGGTGTTAGCTAGGAT 84 1383 741380 N/A N/A 82332 82348GTCTTTTTACATTATAA 31 1384 741381 N/A N/A 82557 82573 ATAACCACTATTCAATG 2 1385 741382 N/A N/A 82783 82799 AAAAATCACATTTGGCA 48 1386 741383 N/AN/A 83008 83024 TTCTTTCACCTTATGAG 21 1387 741384 N/A N/A 83233 83249ATATATGTGTCAGTTCT 22 1388 741385 N/A N/A 83458 83474 GTGTCACTTTTTAAGGT18 1389 741386 N/A N/A 83528 83544 AGAACAATGTCATCTTT 43 1390 741387 N/AN/A 83530 83546 AAAGAACAATGTCATCT 42 1391 741388 N/A N/A 83531 83547GAAAGAACAATGTCATC 25   98 741389 N/A N/A 83532 83548 GGAAAGAACAATGTCAT27 1392 741390 N/A N/A 83534 83550 CAGGAAAGAACAATGTC 47 1393 741391 N/AN/A 83683 83699 CACAGGTATACACACTT 42 1394 741392 N/A N/A 83908 83924GTACAAAATCTGCATAT  3 1395 741393 N/A N/A 84133 84149 ATAGGTATTTTATGCAT58 1396 741394 N/A N/A 84616 84632 CAAATTATGCATTTGTT  0 1397

Example 5: Effect of 5-10-5 MOE Gapmers with Mixed InternucleosideLinkages on Human SNCA In Vitro, Single Dose

Modified oligonucleotides complementary to a human SNCA nucleic acidwere designed and tested for their effect on SNCA mRNA in vitro. Themodified oligonucleotides were tested in a series of experiments thathad similar culture conditions.

Cultured SH-SY5Y cells at a density of 20,000 cells per well weretransfected using electroporation with 4,000 nM concentration ofmodified oligonucleotide or no modified oligonucleotide for untreatedcontrols. After approximately 24 hours, RNA was isolated from the cellsand SNCA mRNA levels were measured by quantitative real-time PCR usinghuman primer probe set RTS2621 as described in Example 1. SNCA mRNAlevels were adjusted according to total RNA content, as measured byRIBOGREEN®. Results are presented in the tables below as percentreduction of the amount of SNCA mRNA, relative to untreated controlcells.

The modified oligonucleotides in tables 29-44 are 5-10-5 MOE gapmers.The gapmers are 20 nucleobases in length, wherein the central gapsegment comprises ten 2′-deoxynucleosides and is flanked by wingsegments on both the 5′ end on the 3′ end, each comprising five 2′-MOEnucleosides. The sugar motif for the gapmers is (from 5′ to 3′):eeeeeddddddddddeeeee; wherein ‘d’ represents a 2′-deoxyribose sugar and‘e’ represents a 2′-MOE modified sugar. All cytosine residues throughouteach gapmer are 5-methyl cytosines. The internucleoside linkages aremixed phosphodiester and phosphorothioate linkages. The internucleosidelinkage motif for the gapmers is (from 5′ to 3′): sooosssssssssssooss;wherein ‘o’ represents a phosphodiester internucleoside linkage and ‘s’represents a phosphorothioate internucleoside linkage. “Start Site”indicates the 5′-most nucleoside to which the gapmer is complementary inthe human nucleic acid sequence. “Stop Site” indicates the 3′-mostnucleoside to which the gapmer is complementary in the human nucleicacid sequence.

Each modified oligonucleotide listed in the Tables below iscomplementary to human SNCA nucleic acid sequences SEQ ID NO: 1 or SEQID NO:2, as indicated. ‘N/A’ indicates that the modified oligonucleotideis not complementary to that particular nucleic acid with 100%complementarity. A value of 0% reduction indicates that the compound hadno effect or increased mRNA concentrations in the cell. As shown below,modified oligonucleotides complementary to human SNCA reduced the amountof human SNCA mRNA.

TABLE 29 Percent reduction of human SNCA mRNA with 5-10-5 MOEgapmers with mixed internucleoside linkages % Com- SEQ ID SEQ SEQ IDSEQ ID Re- SEQ pound No: 1 No: 1 No: 2 No: 2 duc- ID No start stop startstop Sequence (5′ to 3′) tion NO 693413 19 38 4687 4706AATTCCTTTACACCACACTG 53 28 693416 39 58 4707 4726 GAATACATCCATGGCTAATG38 29 740410 240 256 4691 4707 GAATTCCTTTACACCAC 72 33 741410 N/A N/A87946 87962 GTAAGTTGTGACCATGC 64 402 762833 233 252 N/A N/ATCCTTTACACCACACTGTCG 50 1635 762834 234 253 N/A N/A TTCCTTTACACCACACTGTC49 1636 762835 235 254 N/A N/A ATTCCTTTACACCACACTGT 39 1637 762836 237256 4688 4707 GAATTCCTTTACACCACACT 50 1638 762837 238 257 4689 4708TGAATTCCTTTACACCACAC 45 1639 762838 239 258 4690 4709ATGAATTCCTTTACACCACA 47 1640 762839 240 259 4691 4710AATGAATTCCTTTACACCAC 51 1641 762840 241 260 4692 4711TAATGAATTCCTTTACACCA 41 1642 762841 242 261 4693 4712CTAATGAATTCCTTTACACC 44 1643 762842 243 262 4694 4713GCTAATGAATTCCTTTACAC 51 1644 762843 244 263 4695 4714GGCTAATGAATTCCTTTACA 50 1645 762844 252 271 4703 4722ACATCCATGGCTAATGAATT 45 1646 762845 253 272 4704 4723TACATCCATGGCTAATGAAT 32 1647 762846 254 273 4705 4724ATACATCCATGGCTAATGAA 34 1648 762847 255 274 4706 4725AATACATCCATGGCTAATGA 32 1649 762848 257 276 4708 4727TGAATACATCCATGGCTAAT 47 1650 762849 258 277 4709 4728ATGAATACATCCATGGCTAA 43 1651 762850 259 278 4710 4729CATGAATACATCCATGGCTA 41 1652 762851 260 279 4711 4730TCATGAATACATCCATGGCT 61 1653 762852 261 280 4712 4731TTCATGAATACATCCATGGC 48 1654 762853 262 281 4713 4732TTTCATGAATACATCCATGG 42 1655 762854 263 282 4714 4733CTTTCATGAATACATCCATG 56 1656 762855 265 284 4716 4735TCCTTTCATGAATACATCCA 61 1657 762856 49 68 4717 4736 GTCCTTTCATGAATACATCC52 1658 762857 267 286 4718 4737 AGTCCTTTCATGAATACATC 38 1659 762858 268287 4719 4738 AAGTCCTTTCATGAATACAT 60 1660 762859 269 288 4720 4739AAAGTCCTTTCATGAATACA 51 1661 762860 270 289 4721 4740GAAAGTCCTTTCATGAATAC 55 1662 762861 271 290 4722 4741TGAAAGTCCTTTCATGAATA 44 1663 762862 272 291 4723 4742TTGAAAGTCCTTTCATGAAT 34 1664 762863 56 75 4724 4743 TTTGAAAGTCCTTTCATGAA28 1665 762864 432 451 17999 18018 ACTTGCTCTTTGGTCTTCTC 36 1666 762865433 452 18000 18019 CACTTGCTCTTTGGTCTTCT 36 1667 762866 434 453 1800118020 TCACTTGCTCTTTGGTCTTC 40 1668 762867 435 454 18002 18021GTCACTTGCTCTTTGGTCTT 50 1669 762868 436 455 18003 18022TGTCACTTGCTCTTTGGTCT 45 1670 762869 437 456 18004 18023TTGTCACTTGCTCTTTGGTC 36 1671 762870 438 457 18005 18024TTTGTCACTTGCTCTTTGGT 28 1672 762871 439 458 18006 18025ATTTGTCACTTGCTCTTTGG 34 1673 762872 440 459 18007 18026CATTTGTCACTTGCTCTTTG 41 1674 762873 441 460 18008 18027ACATTTGTCACTTGCTCTTT 7 1675 762874 442 461 18009 18028AACATTTGTCACTTGCTCTT 22 1676 762875 N/A N/A 4681 4700TTTACACCACACTGGAAAAC 13 1677 762876 N/A N/A 4682 4701CTTTACACCACACTGGAAAA 22 1678 762877 N/A N/A 4683 4702CCTTTACACCACACTGGAAA 44 1679 762878 N/A N/A 4684 4703TCCTTTACACCACACTGGAA 44 1680 762879 N/A N/A 4685 4704TTCCTTTACACCACACTGGA 45 1681 762880 N/A N/A 4686 4705ATTCCTTTACACCACACTGG 59 1682 762881 N/A N/A 18150 18169TATAAATGTAACACAAAACG 0 1683 762882 N/A N/A 18255 18274GTAGCACTTTTTCACAAGGG 67 1684 762883 N/A N/A 18349 18368CTTTCTTCCAGAAATTGAAA 49 1685 762884 N/A N/A 18442 18461AAATTCCAAGACTTACAATT 28 1686 762885 N/A N/A 18535 18554AGAGATGATGTCACTATAAA 50 1687 762886 N/A N/A 18628 18647TCTCTGGTTGGTATGTATTT 62 1688 762887 N/A N/A 18721 18740TATCTTTGGTATAATCTTAT 41 1689 762888 N/A N/A 18814 18833TTATTTTGCTGTTGTAGTGG 35 1690 762889 N/A N/A 18907 18926GGCAGGCCTCCCCAAGAACG 35 1691 762890 N/A N/A 19176 19195GGCAAAGAAAGGAAAAAGAA 5 1692 762891 N/A N/A 19269 19288ATGGTGCCTACATTCTAGAA 69 1693 762892 N/A N/A 19368 19387CTTAATTTAATAAATGTTTG 7 1694 762893 N/A N/A 19461 19480TTGGATAGCTGAATAGCACT 58 1695 762894 N/A N/A 19556 19575TGAAGTGGAAACAACCCAGA 46 1696 762895 N/A N/A 19627 19646ATTTTTGTTCTGCCTTTTTA 39 1697 762896 N/A N/A 19628 19647TATTTTTGTTCTGCCTTTTT 49 1698 762897 N/A N/A 19629 19648ATATTTTTGTTCTGCCTTTT 34 1699 762898 N/A N/A 19630 19649GATATTTTTGTTCTGCCTTT 57 1700 762899 N/A N/A 19631 19650AGATATTTTTGTTCTGCCTT 60 1701 762900 N/A N/A 19632 19651CAGATATTTTTGTTCTGCCT 74 1702 762901 N/A N/A 19633 19652ACAGATATTTTTGTTCTGCC 70 1703 762902 N/A N/A 19634 19653CACAGATATTTTTGTTCTGC 48 1704 762903 N/A N/A 19635 19654TCACAGATATTTTTGTTCTG 58 1705 762904 N/A N/A 19636 19655ATCACAGATATTTTTGTTCT 55 1706 762905 N/A N/A 19637 19656TATCACAGATATTTTTGTTC 56 1707 762906 N/A N/A 19649 19668TAAATCTAAATATATCACAG 15 1708 762907 N/A N/A 19742 19761AACATTAGCTGAAGAACTTC 36 1709 762908 N/A N/A 19835 19854AACCAGGAATTAATATAATT 33 1710

TABLE 30 Percent reduction of human SNCA mRNA with 5-10-5 MOEgapmers with mixed internucleoside linkages % Com- SEQ ID SEQ ID SEQ IDSEQ ID Re- SEQ pound No: 1 No: 1 No: 2 No: 2 duc- ID No start stop startstop Sequence (5′ to 3′) tion NO 740410 240 256 4691 4707GAATTCCTTTACACCAC 60 33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC 72402 762837 238 257 4689 4708 TGAATTCCTTTACACCACAC 49 1639 762909 N/A N/A19928 19947 CTGATTAATTGCCTGTGTAC 48 1711 762910 N/A N/A 20026 20045ATTAAGCTCTTTGATGTGCG 65 1712 762911 N/A N/A 20119 20138TCTTGGACTACCCACTTCCT 45 1713 762912 N/A N/A 20212 20231AAGGAAGGTAAGTTTTGAGG 0 1714 762913 N/A N/A 20305 20324AGACGGTACATGTTTCCCTG 47 1715 762914 N/A N/A 20332 20351TGTATGCCTTAAATGCAGGT 69 1716 762915 N/A N/A 20398 20417TGGGTGGAAAGCAAACCCAG 16 1717 762916 N/A N/A 20491 20510CTACCTATAAGGGAAATATC 20 1718 762917 N/A N/A 20584 20603TCACTCTCAGTCCAATGTTT 55 1719 762918 N/A N/A 20677 20696GAGCTTCCTCATTTTATGAG 41 1720 762919 N/A N/A 20770 20789ACCACAATTGCAGACATTTA 41 1721 762920 N/A N/A 20871 20890TCAAAGTTTAAAAAATGAAA 6 1722 762921 N/A N/A 20964 20983TCTTGAATGATGAATGAGTG 59 1723 762922 N/A N/A 20979 20998TGAGCAAACATTCTTTCTTG 59 1724 762923 N/A N/A 20980 20999GTGAGCAAACATTCTTTCTT 62 1725 762924 N/A N/A 20981 21000TGTGAGCAAACATTCTTTCT 74 1726 762925 N/A N/A 20982 21001GTGTGAGCAAACATTCTTTC 52 1727 762926 N/A N/A 20983 21002AGTGTGAGCAAACATTCTTT 75 1728 762927 N/A N/A 20984 21003CAGTGTGAGCAAACATTCTT 50 1729 762928 N/A N/A 20985 21004ACAGTGTGAGCAAACATTCT 57 1730 762929 N/A N/A 20986 21005CACAGTGTGAGCAAACATTC 63 1731 762929 N/A N/A 27411 27430CACAGTGTGAGCAAACATTC 63 1731 762930 N/A N/A 20987 21006GCACAGTGTGAGCAAACATT 75 1732 762931 N/A N/A 20988 21007GGCACAGTGTGAGCAAACAT 51 1733 762932 N/A N/A 20989 21008TGGCACAGTGTGAGCAAACA 72 1734 762933 N/A N/A 20993 21012TTTCTGGCACAGTGTGAGCA 43 1735 762934 N/A N/A 20994 21013GTTTCTGGCACAGTGTGAGC 59 1736 762935 N/A N/A 20995 21014AGTTTCTGGCACAGTGTGAG 53 1737 762936 N/A N/A 20996 21015AAGTTTCTGGCACAGTGTGA 44 1738 762937 N/A N/A 20997 21016CAAGTTTCTGGCACAGTGTG 50 1739 762938 N/A N/A 20998 21017CCAAGTTTCTGGCACAGTGT 42 1740 762939 N/A N/A 20999 21018TCCAAGTTTCTGGCACAGTG 51 1741 762940 N/A N/A 21000 21019CTCCAAGTTTCTGGCACAGT 40 1742 762941 N/A N/A 21001 21020CCTCCAAGTTTCTGGCACAG 51 1743 762942 N/A N/A 21002 21021TCCTCCAAGTTTCTGGCACA 57 1744 762943 N/A N/A 21003 21022TTCCTCCAAGTTTCTGGCAC 32 1745 762944 N/A N/A 21057 21076ATTAATCCACTTCTACAAGC 30 1746 762945 N/A N/A 21150 21169GAGGGTGATGGACCAGATAC 51 1747 762946 N/A N/A 21218 21237CAGAATTATGTCATTTAATT 40 1748 762947 N/A N/A 21219 21238TCAGAATTATGTCATTTAAT 58 1749 762948 N/A N/A 21220 21239TTCAGAATTATGTCATTTAA 59 1750 762949 N/A N/A 21221 21240GTTCAGAATTATGTCATTTA 56 1751 762950 N/A N/A 21222 21241TGTTCAGAATTATGTCATTT 66 1752 762951 N/A N/A 21223 21242TTGTTCAGAATTATGTCATT 61 1753 762952 N/A N/A 21224 21243GTTGTTCAGAATTATGTCAT 68 1754 762953 N/A N/A 21225 21244GGTTGTTCAGAATTATGTCA 75 1755 762954 N/A N/A 21226 21245TGGTTGTTCAGAATTATGTC 51 1756 762955 N/A N/A 21227 21246TTGGTTGTTCAGAATTATGT 67 1757 762956 N/A N/A 21228 21247ATTGGTTGTTCAGAATTATG 57 1758 762957 N/A N/A 21243 21262ATTTACTCTCGATTTATTGG 65 1759 762958 N/A N/A 21336 21355TCATTTGTCCTTTAACTAGT 55 1760 762959 N/A N/A 21429 21448TAAAAATATGGATCAAAAGA 0 1761 762960 N/A N/A 21522 21541TGTGCACTTTTAACCTGTTT 69 1762 762961 N/A N/A 21616 21635TTAGAACAAGCAGATCTTTC 63 1763 762962 N/A N/A 21709 21728ATAGACCAAGTGTTCTAGTG 68 1764 762963 N/A N/A 21802 21821GAGCATTCCATGTGGCATGA 62 1765 762964 N/A N/A 21895 21914CAGGCAGTGGCTTCACAGTT 40 1766 762965 N/A N/A 21993 22012TTTCAAGCTTATTTCTTGCG 69 1767 762966 N/A N/A 22086 22105AAATGGCATTGCTTAGGAAC 39 1768 762967 N/A N/A 22179 22198AAGTCAGGATTATTACAGAA 51 1769 762968 N/A N/A 22273 22292GATATTATATTCACAATGTC 34 1770 762969 N/A N/A 22366 22385GGTCCATAACACTCAAGTGT 79 1771 762970 N/A N/A 22448 22467GACCTGTGCTTGTTTGTGAA 60 1772 762971 N/A N/A 22449 22468TGACCTGTGCTTGTTTGTGA 58 1773 762972 N/A N/A 22450 22469GTGACCTGTGCTTGTTTGTG 56 1774 762973 N/A N/A 22451 22470TGTGACCTGTGCTTGTTTGT 48 1775 762974 N/A N/A 22452 22471CTGTGACCTGTGCTTGTTTG 61 1776 762975 N/A N/A 22453 22472CCTGTGACCTGTGCTTGTTT 47 1777 762976 N/A N/A 22454 22473GCCTGTGACCTGTGCTTGTT 50 1778 762977 N/A N/A 22455 22474TGCCTGTGACCTGTGCTTGT 54 1779 762978 N/A N/A 22456 22475TTGCCTGTGACCTGTGCTTG 59 1780 762979 N/A N/A 22457 22476GTTGCCTGTGACCTGTGCTT 68 1781 762980 N/A N/A 22458 22477TGTTGCCTGTGACCTGTGCT 54 1782 762981 N/A N/A 22459 22478ATGTTGCCTGTGACCTGTGC 40 1783 762982 N/A N/A 22460 22479AATGTTGCCTGTGACCTGTG 28 1784 762983 N/A N/A 22461 22480AAATGTTGCCTGTGACCTGT 49 1785 762984 N/A N/A 22462 22481GAAATGTTGCCTGTGACCTG 30 1786 762985 N/A N/A 22463 22482TGAAATGTTGCCTGTGACCT 49 1787

TABLE 31 Percent reduction of human SNCA mRNA with 5-10-5 MOEgapmers with mixed internucleoside linkages % Com- SEQ ID SEQ ID SEQ IDSEQ ID Re- SEQ pound No: 1 No: 1 No: 2 No: 2 duc- ID No start stop startstop Sequence (5′ to 3′) tion NO 740410 240 256 4691 4707GAATTCCTTTACACCAC 83 33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC 94402 762837 238 257 4689 4708 TGAATTCCTTTACACCACAC 52 1639 762986 N/A N/A22464 22483 CTGAAATGTTGCCTGTGACC 58 1788 762987 N/A N/A 22552 22571CTCCAGTCCTGACATCTCTT 77 1789 762988 N/A N/A 22645 22664ACAAGAACCAAACTTTTAAT 57 1790 762989 N/A N/A 22738 22757CAAATCAGGCAATTCATTGT 57 1791 762990 N/A N/A 22831 22850AAATCAATCTTAAATTTTTC 0 1792 762991 N/A N/A 22924 22943TTTATGTACCATTAGTGGGC 63 1793 762992 N/A N/A 23017 23036CATTAGAATTCACTATTCAT 54 1794 762993 N/A N/A 23110 23129TTAATGAAAACATAGCAGTA 37 1795 762994 N/A N/A 23203 23222AAGGCAGGAGCCACCCATAT 56 1796 762995 N/A N/A 23279 23298TTCTCAGAGACCACAGCTGC 70 1797 762996 N/A N/A 23280 23299GTTCTCAGAGACCACAGCTG 70 1798 762997 N/A N/A 23281 23300TGTTCTCAGAGACCACAGCT 64 1799 762998 N/A N/A 23282 23301TTGTTCTCAGAGACCACAGC 68 1800 762999 N/A N/A 23283 23302ATTGTTCTCAGAGACCACAG 67 1801 763000 N/A N/A 23284 23303TATTGTTCTCAGAGACCACA 57 1802 763001 N/A N/A 23285 23304ATATTGTTCTCAGAGACCAC 71 1803 763002 N/A N/A 23286 23305CATATTGTTCTCAGAGACCA 81 1804 763003 N/A N/A 23287 23306CCATATTGTTCTCAGAGACC 67 1805 763004 N/A N/A 23288 23307ACCATATTGTTCTCAGAGAC 69 1806 763005 N/A N/A 23289 23308AACCATATTGTTCTCAGAGA 65 1807 763006 N/A N/A 23290 23309AAACCATATTGTTCTCAGAG 68 1808 763007 N/A N/A 23291 23310CAAACCATATTGTTCTCAGA 64 1809 763008 N/A N/A 23296 23315TGTAACAAACCATATTGTTC 73 1810 763009 N/A N/A 23389 23408CAAAAACACAATTTAATGTA 14 1811 763010 N/A N/A 23482 23501GATTTGGGTGGAAGTATTTG 47 1812 763011 N/A N/A 23575 23594CGCAATCAGTTCTTTGAATA 73 1813 763012 N/A N/A 23668 23687CAAATATGATTTAAACCTAT 4 1814 763013 N/A N/A 23761 23780ATGGGTTCACAGAAGTGTGG 65 1815 763014 N/A N/A 23854 23873ACAGTATCTCATTAATGAAA 45 1816 763015 N/A N/A 23948 23967ATAAGTATCTCAAAACATCA 52 1817 763016 N/A N/A 24041 24060AAGATAACCATATGATGATG 42 1818 763017 N/A N/A 24160 24179GTAAGATGAGTAAGTCTAAA 59 1819 763018 N/A N/A 24253 24272ACATATAAGTGCTATTTTTC 42 1820 763019 N/A N/A 24346 24365AGGGACAAACAGGTTGTTTA 83 1821 763020 N/A N/A 24439 24458AAAGCAAATAGCATCATCAA 44 1822 763021 N/A N/A 24539 24558CTGTACCCTTGAATATCACG 69 1823 763022 N/A N/A 24632 24651ACAATTAAATTGATGAGATG 18 1824 763023 N/A N/A 24731 24750CTTAAAAATCCAAATGTTGT 51 1825 763024 N/A N/A 24825 24844CATTAATAAGAATTAAATGC 6 1826 763025 N/A N/A 24850 24869TTCTTTGCATTAGTATTCAC 53 1827 763026 N/A N/A 24851 24870ATTCTTTGCATTAGTATTCA 48 1828 763027 N/A N/A 24852 24871TATTCTTTGCATTAGTATTC 48 1829 763028 N/A N/A 24853 24872GTATTCTTTGCATTAGTATT 60 1830 763029 N/A N/A 24854 24873AGTATTCTTTGCATTAGTAT 72 1831 763030 N/A N/A 24855 24874CAGTATTCTTTGCATTAGTA 69 1832 763031 N/A N/A 24856 24875TCAGTATTCTTTGCATTAGT 70 1833 763032 N/A N/A 24857 24876CTCAGTATTCTTTGCATTAG 77 1834 763033 N/A N/A 24858 24877GCTCAGTATTCTTTGCATTA 79 1835 763034 N/A N/A 24859 24878GGCTCAGTATTCTTTGCATT 69 1836 763035 N/A N/A 24860 24879TGGCTCAGTATTCTTTGCAT 77 1837 763036 N/A N/A 24918 24937TCCATTTTTTCACTTACTTG 75 1838 763037 N/A N/A 25011 25030TTAGATTTATCATATTGTTG 50 1839 763038 N/A N/A 25104 25123TTAAAATCTATTTGATTTCA 32 1840 763039 N/A N/A 25198 25217CCAAATAGAAAAAAAGTGTG 18 1841 763040 N/A N/A 25291 25310CTGTATGTACAACCTCAGAA 82 1842 763041 N/A N/A 25384 25403CCTGACATAAGTAGGAAGCA 63 1843 763042 N/A N/A 25477 25496CCTACTTTAGATATGTCATA 69 1844 763043 N/A N/A 25570 25589TGTTAGTATACCTTTGTAGG 72 1845 763044 N/A N/A 25663 25682GAGGGCCAGCTGGCCATCAT 15 1846 763045 N/A N/A 25756 25775GAATTCAAGCCCATGCCCTC 44 1847 763046 N/A N/A 25854 25873CAACATTTTTATTTCACAGA 54 1848 763047 N/A N/A 25947 25966GTTGCCAGGGATCTGGCAAC 15 1849 763048 N/A N/A 26040 26059TGTCTGCATTATCTTATTTC 67 1850 763049 N/A N/A 26133 26152TGTGATCATGTATCGACACA 78 1851 763050 N/A N/A 26226 26245AACGGCATGTTCAGTGATGC 82 1852 763051 N/A N/A 26319 26338ATTATACCATGTGCATAATA 54 1853 763052 N/A N/A 26412 26431GCCTTTGAGATTTGCTTCAG 91 1854 763053 N/A N/A 26505 26524TTTTACTGAACACCTAGAAC 54 1855 763054 N/A N/A 26598 26617TTCATCTAGGACCTGCAATC 39 1856 763055 N/A N/A 26691 26710TTGGTGTTGTCCCAAGAAAT 63 1857 763056 N/A N/A 26784 26803CTGCAATCTACTTAGACCTG 71 1858 763057 N/A N/A 26877 26896AGGAAAAACTCTGCCCTCCT 46 1859 763058 N/A N/A 26978 26997CAAATGAACTTGGGAGGAGG 14 1860 763059 N/A N/A 27071 27090AGTGCAGGATGAAACCAGAC 76 1861 763060 N/A N/A 27164 27183TGAAGTATTAGAGAGGATCA 46 1862 763061 N/A N/A 27257 27276CTCGGACGGAAGTGAAGGCA 57 1863 763062 N/A N/A 27350 27369GCTCACTTCCTGTCACCCCC 49 1864

TABLE 32 Percent reduction of human SNCA mRNA with 5-10-5 MOE gapmerswith mixed internucleoside linkages % SEQ ID SEQ ID SEQ ID SEQ ID Re-SEQ Compound No: 1 No: 1 No: 2 No: 2 duc- ID No start stop start stopSequence (5′ to 3′) tion NO 740410 240 256 4691 4707 GAATTCCTTTACACCAC76 33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC 64 402 762837 238 2574689 4708 TGAATTCCTTTACACCACAC 61 1639 763063 N/A N/A 27443 27462TAGGATGCAGCCTGAGGAGC 29 1865 763064 N/A N/A 27536 27555CATTAGAGTTTGTCTCTGGT 58 1866 763065 N/A N/A 27629 27648GACCCTTTCATTACCTTTCA 81 1867 763066 N/A N/A 27722 27741TCCTAGCCCACATCTTAGTA 19 1868 763067 N/A N/A 27815 27834ATTGTTCTGATTGATGGACA 59 1869 763068 N/A N/A 27908 27927TGCGACTGGTCAGAGCATGC 63 1870 763069 N/A N/A 28001 28020TCTAGCACTATTTTTTTCAA 31 1871 763070 N/A N/A 28094 28113TTAATAATTATTCTACAACA 0 1872 763071 N/A N/A 28192 28211CACATACAGGTTTTTAAAAA 21 1873 763072 N/A N/A 28231 28250CCTTGTGCATTTATTCCACG 84 1874 763073 N/A N/A 28232 28251ACCTTGTGCATTTATTCCAC 67 1875 763074 N/A N/A 28233 28252TACCTTGTGCATTTATTCCA 64 1876 763075 N/A N/A 28234 28253GTACCTTGTGCATTTATTCC 70 1877 763076 N/A N/A 28235 28254AGTACCTTGTGCATTTATTC 68 1878 763077 N/A N/A 28236 28255GAGTACCTTGTGCATTTATT 67 1879 763078 N/A N/A 28286 28305CGAAGAATTACCCAGCCCAA 28 1880 763079 N/A N/A 28379 28398GTGCTTGTTGCCATGCTGGG 78 1881 763080 N/A N/A 28451 28470TGTAATCTAGGACCCAGTAA 56 1882 763081 N/A N/A 28452 28471CTGTAATCTAGGACCCAGTA 70 1883 763082 N/A N/A 28453 28472ACTGTAATCTAGGACCCAGT 63 1884 763083 N/A N/A 28454 28473GACTGTAATCTAGGACCCAG 69 1885 763084 N/A N/A 28455 28474AGACTGTAATCTAGGACCCA 72 1886 763085 N/A N/A 28456 28475CAGACTGTAATCTAGGACCC 70 1887 763086 N/A N/A 28457 28476CCAGACTGTAATCTAGGACC 64 1888 763087 N/A N/A 28458 28477TCCAGACTGTAATCTAGGAC 92 1889 763088 N/A N/A 28459 28478ATCCAGACTGTAATCTAGGA 83 1890 763089 N/A N/A 28460 28479AATCCAGACTGTAATCTAGG 51 1891 763090 N/A N/A 28461 28480TAATCCAGACTGTAATCTAG 49 1892 763091 N/A N/A 28472 28491AAGGAACGCAATAATCCAGA 47 1893 763092 N/A N/A 28565 28584ACCAGTGCGGAATATTGTAA 64 1894 763093 N/A N/A 28669 28688ATCAGTCGAATGAATGTACG 36 1895 763094 N/A N/A 28765 28784CAGATGGATGGGTGGACAAA 52 1896 763095 N/A N/A 29117 29136TTGGCATTGTATTTTTTTTG 60 1897 763096 N/A N/A 29210 29229TAGACTCCTACACATATTAA 32 1898 763097 N/A N/A 29303 29322GATACTTCACTCAGAAAACC 34 1899 763098 N/A N/A 29396 29415AAAATGGTTTGATAGTTGGG 55 1900 763099 N/A N/A 29454 29473TTGTTAATAGTTCTCTGTTT 62 1901 763100 N/A N/A 29455 29474TTTGTTAATAGTTCTCTGTT 45 1902 763101 N/A N/A 29456 29475TTTTGTTAATAGTTCTCTGT 54 1903 763102 N/A N/A 29457 29476TTTTTGTTAATAGTTCTCTG 70 1904 763103 N/A N/A 29489 29508GGATACCATACAACCAATTA 57 1905 763104 N/A N/A 29582 29601ACAACTAAATCACTCAATTC 8 1906 763105 N/A N/A 29675 29694CAGAGCCCAAAACATTTATA 33 1907 763106 N/A N/A 29801 29820CAAATGCCTTGATCTTGGAG 42 1908 763107 N/A N/A 29894 29913GAGAACACAGCATTTGGCCC 68 1909 763108 N/A N/A 29997 30016AGAGGTAATAAAGTCACGGG 46 1910 763109 N/A N/A 30090 30109ATATGAAAATGAAAGGATGG 28 1911 763110 N/A N/A 30193 30212TTACAGTTTCCTATATATCG 25 1912 763111 N/A N/A 30287 30306TCATACACAAAATAAACACA 33 1913 763112 N/A N/A 30380 30399GAATAGCAGTATGTACTAAT 40 1914 763113 N/A N/A 30473 30492ACCTTTCAATAAACTGTTAA 33 1915 763114 N/A N/A 30566 30585ATTTTTTCATATATAGTGAG 49 1916 763115 N/A N/A 30659 30678CTGTAACAAATATACATTTT 36 1917 763116 N/A N/A 30752 30771ACCAATTAGTTTCTAATAAG 38 1918 763117 N/A N/A 30885 30904TTATATACACACACAGCTAC 14 1919 763118 N/A N/A 30978 30997CCAAAAATAGAGATCAATGT 31 1920 763119 N/A N/A 31078 31097AAACCACTGGCTAATTTTTT 57 1921 763120 N/A N/A 31171 31190TGAGAGCTATATGGCTGAAA 47 1922 763121 N/A N/A 31264 31283AAAAGCCTTCTTAGTGGTAA 53 1923 763122 N/A N/A 31357 31376ATTACTGTGTTTCAGCAGTT 51 1924 763123 N/A N/A 31450 31469TTAGATATAAAAGGTATGAA 0 1925 763124 N/A N/A 31543 31562CTAGCCTAGGGTGGTAACAG 17 1926 763125 N/A N/A 31636 31655TGCTCAAGAATGGACTAGGT 57 1927 763126 N/A N/A 31729 31748TGCATTTCATTCTATGTATG 62 1928 763127 N/A N/A 31822 31841AGTTGGAGGGTGGCATACAA 25 1929 763128 N/A N/A 31915 31934AACAAACAATTCATTTTCTA 0 1930 763129 N/A N/A 32011 32030GTCTTTTTAAAATTAAAATC 0 1931 763130 N/A N/A 32104 32123TATAATATACAAAATTACTA 0 1932 763131 N/A N/A 32197 32216CAATAAGTAGTGCTGTTATA 51 1933 763132 N/A N/A 32290 32309AGTAGTTTTTAAATCTTCAA 51 1934 763133 N/A N/A 32383 32402GTGGTTCTGCCCATCTGTCC 64 1935 763134 N/A N/A 32476 32495TGTTTTCAAGAGCGATCGGA 58 1936 763135 N/A N/A 32569 32588GAGTAAGTTTAGATATAAAA 36 1937 763136 N/A N/A 32662 32681GCATAAAAGGCAGAGGGAGG 35 1938 763137 N/A N/A 32755 32774GCAACCTTTCTCTCCCTCTC 65 1939 763138 N/A N/A 32848 32867CTGAAGAAATAAATAAAGAA 0 1940 763139 N/A N/A 32941 32960TCAATATTCAGAGATGACTA 27 1941

TABLE 33 Percent reduction of human SNCA mRNA with 5-10-5 MOE gapmerswith mixed internucleoside linkages % SEQ ID SEQ ID SEQ ID SEQ ID Re-SEQ Compound No: 1 No: 1 No: 2 No: 2 duc- ID No start stop start stopSequence (5′ to 3′) tion NO 740410 240 256 4691 4707 GAATTCCTTTACACCAC78 33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC 72 402 762837 238 2574689 4708 TGAATTCCTTTACACCACAC 46 1639 763140 N/A N/A 33034 33053CACACCCGAAATACCACCTG 49 1942 763141 N/A N/A 33127 33146TAAGCTAAAATGGTTTCAAC 45 1943 763142 N/A N/A 33220 33239GGCAAAATTTGCATTGGATG 79 1944 763143 N/A N/A 33313 33332TGAAAAAAGCTATGACCCTC 30 1945 763144 N/A N/A 33406 33425TTACTTCTACTTTTGTGAGG 46 1946 763145 N/A N/A 33499 33518CCAACATTTTAAGGAAGGTA 73 1947 763146 N/A N/A 33592 33611GGTAGAGAACACTTAAGTGA 67 1948 763147 N/A N/A 33689 33708CAAATTTTAAAAGTTAACTT 0 1949 763148 N/A N/A 33785 33804AATTTCTACAAGAAAAATAT 0 1950 763149 N/A N/A 33878 33897CCAGAAAGAAACATTTAGAA 39 1951 763150 N/A N/A 33971 33990GTGTTAACTGGCAATTCCAT 82 1952 763151 N/A N/A 33981 34000GTTCAATGCTGTGTTAACTG 74 1953 763152 N/A N/A 33982 34001AGTTCAATGCTGTGTTAACT 63 1954 763153 N/A N/A 33983 34002AAGTTCAATGCTGTGTTAAC 50 1955 763154 N/A N/A 33984 34003AAAGTTCAATGCTGTGTTAA 38 1956 763155 N/A N/A 33985 34004AAAAGTTCAATGCTGTGTTA 49 1957 763156 N/A N/A 33986 34005AAAAAGTTCAATGCTGTGTT 63 1958 763157 N/A N/A 33987 34006GAAAAAGTTCAATGCTGTGT 56 1959 763158 N/A N/A 33988 34007AGAAAAAGTTCAATGCTGTG 62 1960 763159 N/A N/A 33989 34008AAGAAAAAGTTCAATGCTGT 42 1961 763160 N/A N/A 33990 34009CAAGAAAAAGTTCAATGCTG 52 1962 763161 N/A N/A 33991 34010ACAAGAAAAAGTTCAATGCT 28 1963 763162 N/A N/A 34064 34083TAATATCAGCCAAAGACATT 34 1964 763163 N/A N/A 34157 34176TTGAAAAAAGTATTGACTCT 16 1965 763164 N/A N/A 34250 34269CTGTTAAAATGCATTTCTAG 64 1966 763165 N/A N/A 34383 34402GTATCCCAGCACTGTTGGGA 25 1967 763166 N/A N/A 34476 34495CTGGTTGCTATCTAGGGATC 82 1968 763167 N/A N/A 34569 34588AATAGAACCTAATATAATTT 0 1969 763168 N/A N/A 34662 34681CTTCTATCCTAGAATTCATA 40 1970 763169 N/A N/A 34755 34774ATGGGAATGAGGTGTAAAAG 56 1971 763170 N/A N/A 34848 34867CAGTCTGATAAGGAGAACAA 45 1972 763171 N/A N/A 34941 34960GGATAGAATATCAAGATAAA 38 1973 763172 N/A N/A 35034 35053TCACAGTGTTCTTTTCTCTT 71 1974 763173 N/A N/A 35127 35146TTGTGCTAGAATATGAGATC 0 1975 763174 N/A N/A 35220 35239TCTAGAATTCAAGCCACACC 41 1976 763175 N/A N/A 35313 35332AATAATGATAGTATTTTCCT 15 1977 763176 N/A N/A 35406 35425CCATCACACCTTGCAGATGT 70 1978 763177 N/A N/A 35499 35518ACTTCCTTTAGAGTATACGG 79 1979 763178 N/A N/A 35594 35613GCACTATATAAAATGTAACG 68 1980 763179 N/A N/A 35687 35706GTTGTAATAATAATATTGAC 51 1981 763180 N/A N/A 35780 35799CTGTGACTTTGGTCTATTTG 34 1982 763181 N/A N/A 35873 35892GGATTGTGTAATAGCCTTTA 60 1983 763182 N/A N/A 35966 35985TGACTATCAGTATCTGTTGA 75 1984 763183 N/A N/A 36059 36078AAGTTGACTTGTGACATACA 45 1985 763184 N/A N/A 36152 36171TTCTACCGAAGGAAATATGT 27 1986 763185 N/A N/A 36250 36269AGTCATTTTAATAAGTGTTT 72 1987 763186 N/A N/A 36360 36379ATCTTCCAAAGTTACTGTAC 49 1988 763187 N/A N/A 36453 36472ATTTCCCAGTCTCGGGAACT 17 1989 763188 N/A N/A 36625 36644GCTAATGGTTTTATGTGTTT 73 1990 763189 N/A N/A 36789 36808CATATGGTGATATGGTTAGG 55 1991 763190 N/A N/A 36933 36952TCATTCACCTATTGAGGAAC 50 1992 763191 N/A N/A 37026 37045CTAAGTTTTCTCCATGTGTT 52 1993 763192 N/A N/A 37135 37154GAGCCCCAGGCAATCACTGA 0 1994 763193 N/A N/A 37229 37248GGTCATGTATCCACCATGAC 44 1995 763194 N/A N/A 37322 37341AAATAACATTGATACCTTAT 40 1996 763195 N/A N/A 37415 37434ATTACAGTGCATTCCCATAT 41 1997 763196 N/A N/A 37523 37542GGGTCTTGACTTCCCAAAGT 74 1998 763197 N/A N/A 37649 37668TCTTTTATTTCTTCTGTTCT 34 1999 763198 N/A N/A 37785 37804CGCATCTGTCTTTCTTTTCT 31 2000 763199 N/A N/A 37878 37897GTAATCTCACCCTACTGCAA 4 2001 763200 N/A N/A 37971 37990TATCTAGACTGAGCTTTACA 39 2002 763201 N/A N/A 38064 38083CATTCACATATTTGGATTCT 60 2003 763202 N/A N/A 38157 38176TGAAACATTAACTGCTTTAT 51 2004 763203 N/A N/A 38250 38269ACAATGCTATGTGGAAGTTA 44 2005 763204 N/A N/A 38343 38362CCTCAGTGCTAGCGAAGGAC 67 2006 763205 N/A N/A 38436 38455AATTTACAATCTACACAGGC 53 2007 763206 N/A N/A 38529 38548TCAATTCTTGAGGCCAATTG 27 2008 763207 N/A N/A 38622 38641TCAGTATTTCATTGTCATAC 89 2009 763208 N/A N/A 38715 38734GTTAGTGGAATTGTAAAATA 54 2010 763209 N/A N/A 38808 38827CTAGTTATAAAAAACAAGAT 34 2011 763210 N/A N/A 38901 38920TTGGCCCCAATCATTGGAAT 43 2012 763211 N/A N/A 38910 38929TTCTCTGTATTGGCCCCAAT 52 2013 763212 N/A N/A 38911 38930TTTCTCTGTATTGGCCCCAA 59 2014 763213 N/A N/A 38912 38931TTTTCTCTGTATTGGCCCCA 51 2015 763214 N/A N/A 38913 38932GTTTTCTCTGTATTGGCCCC 60 2016 763215 N/A N/A 38914 38933TGTTTTCTCTGTATTGGCCC 70 2017 763216 N/A N/A 38915 38934ATGTTTTCTCTGTATTGGCC 74 2018

TABLE 34 Percent reduction of human SNCA mRNA with 5-10-5 MOE gapmerswith mixed internucleoside linkages % SEQ ID SEQ ID SEQ ID SEQ ID Re-SEQ Compound No: 1 No: 1 No: 2 No: 2 duc- ID No start stop start stopSequence (5′ to 3′) tion NO 740410 240 256 4691 4707 GAATTCCTTTACACCAC67 33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC 95 402 762837 238 2574689 4708 TGAATTCCTTTACACCACAC 47 1639 763217 N/A N/A 38916 38935GATGTTTTCTCTGTATTGGC 54 2019 763218 N/A N/A 38917 38936AGATGTTTTCTCTGTATTGG 61 2020 763219 N/A N/A 38918 38937GAGATGTTTTCTCTGTATTG 61 2021 763220 N/A N/A 38919 38938TGAGATGTTTTCTCTGTATT 48 2022 763221 N/A N/A 38920 38939TTGAGATGTTTTCTCTGTAT 52 2023 763222 N/A N/A 38994 39013TTTTCAGCAGGAGTTATAAT 43 2024 763223 N/A N/A 39087 39106TATTCCGTGTGTTTTTCCTA 31 2025 763224 N/A N/A 39180 39199TTTTCTGATAAATGGTAATC 12 2026 763225 N/A N/A 39273 39292CAGGTGGTATCAGTCCAAAG 69 2027 763226 N/A N/A 39366 39385CACCACAAAGAGGAAACAGG 41 2028 763227 N/A N/A 39459 39478AATGTTCCCTGGGAGCACAA 58 2029 763228 N/A N/A 39555 39574ATGTCCTGTGCATTGTAGAT 64 2030 763229 N/A N/A 39585 39604TTTTTTAACTGGATACTTTG 27 2031 763230 N/A N/A 39586 39605ATTTTTTAACTGGATACTTT 26 2032 763231 N/A N/A 39587 39606CATTTTTTAACTGGATACTT 44 2033 763232 N/A N/A 39588 39607ACATTTTTTAACTGGATACT 48 2034 763233 N/A N/A 39589 39608GACATTTTTTAACTGGATAC 66 2035 763234 N/A N/A 39590 39609TGACATTTTTTAACTGGATA 55 2036 763235 N/A N/A 39591 39610ATGACATTTTTTAACTGGAT 51 2037 763236 N/A N/A 39592 39611AATGACATTTTTTAACTGGA 64 2038 763237 N/A N/A 39593 39612TAATGACATTTTTTAACTGG 38 2039 763238 N/A N/A 39594 39613GTAATGACATTTTTTAACTG 53 2040 763239 N/A N/A 39595 39614AGTAATGACATTTTTTAACT 38 2041 763240 N/A N/A 39648 39667GTGCCTGGAGAAGATGAATT 44 2042 763241 N/A N/A 39741 39760CTTTTCCTATTTGGTATTTG 53 2043 763242 N/A N/A 39834 39853TTGCTAAATATTACTCACTC 40 2044 763243 N/A N/A 39927 39946ACCAGACTGACTGTAATATG 60 2045 763244 N/A N/A 40020 40039AAGTGAAAGCATTAGAGGAT 54 2046 763245 N/A N/A 40113 40132TGGTGTGTGCAAACATGTAT 27 2047 763246 N/A N/A 40206 40225TTGTGAGAAAGTTTTTATGG 17 2048 763247 N/A N/A 40299 40318ATAAATAGTCATAAGACTAT 5 2049 763248 N/A N/A 40392 40411AGTGTGATATCTAAATAAAA 11 2050 763249 N/A N/A 40485 40504AATTCTGGTGCCAATGGTGA 70 2051 763250 N/A N/A 40578 40597ATCATCTTATGGCTAAATTT 42 2052 763251 N/A N/A 40671 40690ATCTAGGCATGAGTTGTGTC 39 2053 763252 N/A N/A 40775 40794CGTTTGAATGAAAAATGACG 37 2054 763253 N/A N/A 40868 40887ATTAGAACGAGGATGGAGAA 32 2055 763254 N/A N/A 40961 40980AGAGAATTCACATGATAGAT 44 2056 763255 N/A N/A 41054 41073TAAGAAAGAATTTTAGGCAT 35 2057 763256 N/A N/A 41147 41166GCAGGAGCAACACAGTGAAC 40 2058 763257 N/A N/A 41241 41260GATCAACAGGAAACATTTAT 45 2059 763258 N/A N/A 41334 41353TACCCCTATATCTCAACTCA 43 2060 763259 N/A N/A 41427 41446AATGTTATAGTTTCTACATG 34 2061 763260 N/A N/A 41521 41540CCAATTATGTAATTTTAAAT 0 2062 763261 N/A N/A 41619 41638TCTCATTCAAAACCATCCTG 59 2063 763262 N/A N/A 41740 41759TAATTGTCTTGAGCCATGCA 48 2064 763263 N/A N/A 41833 41852GCTTATAGTACACATTAACT 56 2065 763264 N/A N/A 41933 41952GCCCTCTCTCATTACCGTCG 44 2066 763265 N/A N/A 42026 42045AATACAAATTAGTTGAGTTA 22 2067 763266 N/A N/A 42119 42138ATACCACATACTCATTTTAA 46 2068 763267 N/A N/A 42212 42231TAGTTACATGTAGAATGCAT 41 2069 763268 N/A N/A 42305 42324TCTGGGATACAAGGTGTACC 54 2070 763269 N/A N/A 42398 42417CTTCATGGGAAGAAAAGCTA 33 2071 763270 N/A N/A 42491 42510ACAGAAGTACAGCATGTAAG 51 2072 763271 N/A N/A 42598 42617TATTAAGAGTAATGCTATCG 48 2073 763272 N/A N/A 42691 42710AGTAGTCCATTCCATTTTTG 76 2074 763273 N/A N/A 42785 42804ATTTGTCTTTTCTGGAATTA 47 2075 763274 N/A N/A 42878 42897AATTCTAACACCATCTTGGA 22 2076 763275 N/A N/A 42971 42990CTACATTGTGGTTTTTCCTT 31 2077 763276 N/A N/A 43064 43083GGAAGCCAAGACTTTCTTGT 55 2078 763277 N/A N/A 43157 43176GACTGGCCTCCAGCCAATGA 45 2079 763278 N/A N/A 43250 43269ACCTCTTGGATCTTTTCTCT 41 2080 763279 N/A N/A 43343 43362TTCCCCAATTTTCCTTTGTG 37 2081 763280 N/A N/A 43436 43455CACTCATTTTAAATGTACAT 49 2082 763281 N/A N/A 43529 43548GTCTTAGGTTTATGTTCATG 73 2083 763282 N/A N/A 43622 43641AATGTCACAAGACTTCATCT 59 2084 763283 N/A N/A 43715 43734CCCCTTGAAAATGTATGTTA 47 2085 763284 N/A N/A 43808 43827GACCTCTTAATGTTTCTTTG 53 2086 763285 N/A N/A 43901 43920AGATCAGATCATAATCAATA 42 2087 763286 N/A N/A 43994 44013ACTAGAACTGAGGGACAAGG 13 2088 763287 N/A N/A 44376 44395ATTTTGGGCTGGAAGCAGTG 6 2089 763288 N/A N/A 44469 44488GGCAGGAACAACTCTGTCAG 62 2090 763289 N/A N/A 44574 44593AGCACCAACCAACCAGAGGG 51 2091 763290 N/A N/A 44667 44686CCCTGTCAAATTTTAGAAAT 25 2092 763291 N/A N/A 44826 44845TTGGTCTAACTGTGTTGCCC 65 2093 763292 N/A N/A 45028 45047GAGGATTCACTAATTTTTTT 43 2094 763293 N/A N/A 45121 45140AAAACAAAAGAGAAGCAACC 40 2095

TABLE 35 Percent reduction of human SNCA mRNA with 5-10-5 MOE gapmerswith mixed internucleoside linkages % SEQ ID SEQ ID SEQ ID SEQ ID Re-SEQ Compound No: 1 No: 1 No: 2 No: 2 duc- ID No start stop start stopSequence (5′ to 3′) tion NO 740410 240 256 4691 4707 GAATTCCTTTACACCAC62 33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC 72 402 762837 238 2574689 4708 TGAATTCCTTTACACCACAC 34 1639 763294 N/A N/A 45214 45233GTCTCTAGTTTTCCTAAAAT 53 2096 763295 N/A N/A 45307 45326AGGATACTCAATCTCTTAAT 63 2097 763296 N/A N/A 45400 45419CGAATAGAAAAATTTAACTT 0 2098 763297 N/A N/A 45493 45512CTTCAATTAATATTCCAAGA 54 2099 763298 N/A N/A 45586 45605CACTGTGGATGAAGGTTACT 47 2100 763299 N/A N/A 45679 45698GGACTACTTGATGTCTAGAT 68 2101 763300 N/A N/A 45773 45792TTTAATAATACAGTATTATT 0 2102 763301 N/A N/A 45866 45885AACCTACAGAGAGTGGACTT 34 2103 763302 N/A N/A 45959 45978TTTATTTCCCACATAAAATT 4 2104 763303 N/A N/A 46052 46071TACTTAAGAGAAAAAATAGT 0 2105 763304 N/A N/A 46145 46164CCAAGTAAATAGTATTTTGG 28 2106 763305 N/A N/A 46155 46174TTCCATGAAGCCAAGTAAAT 63 2107 763306 N/A N/A 46156 46175TTTCCATGAAGCCAAGTAAA 44 2108 763307 N/A N/A 46157 46176ATTTCCATGAAGCCAAGTAA 48 2109 763308 N/A N/A 46158 46177GATTTCCATGAAGCCAAGTA 50 2110 763309 N/A N/A 46159 46178AGATTTCCATGAAGCCAAGT 66 2111 763310 N/A N/A 46160 46179GAGATTTCCATGAAGCCAAG 50 2112 763311 N/A N/A 46161 46180AGAGATTTCCATGAAGCCAA 34 2113 763312 N/A N/A 46162 46181GAGAGATTTCCATGAAGCCA 65 2114 763313 N/A N/A 46163 46182AGAGAGATTTCCATGAAGCC 41 2115 763314 N/A N/A 46164 46183GAGAGAGATTTCCATGAAGC 61 2116 763315 N/A N/A 46165 46184TGAGAGAGATTTCCATGAAG 30 2117 763316 N/A N/A 46166 46185GTGAGAGAGATTTCCATGAA 47 2118 763317 N/A N/A 46167 46186AGTGAGAGAGATTTCCATGA 44 2119 763318 N/A N/A 46238 46257GGATTTATGTAACAGGAATA 53 2120 763319 N/A N/A 46331 46350TGATTTAATACATATTTGCA 33 2121 763320 N/A N/A 46424 46443TCCACACTTCCCTCGATACT 18 2122 763321 N/A N/A 46529 46548GTGGTGGTGCCAGCAGTGGG 39 2123 763322 N/A N/A 46622 46641TACCACCCACAGCTGTGCCC 43 2124 763323 N/A N/A 46715 46734TAGAATAGTGCCTGTTTAAA 19 2125 763324 N/A N/A 46808 46827AATTGCCTTTTCTGTTTCTT 48 2126 763325 N/A N/A 46905 46924TACTAGCATAGTGTCTAGCA 43 2127 763326 N/A N/A 46998 47017ATACTCAGACATCTTAAGTC 52 2128 763327 N/A N/A 47093 47112GATGCCTGACACAAAATAGG 54 2129 763328 N/A N/A 47186 47205ATTATATTTTGCCTAACCTC 0 2130 763329 N/A N/A 47279 47298GAGAAAATCTGTCTCCTTGC 27 2131 763330 N/A N/A 47372 47391CATTGTGGGATTGTAAGTCT 31 2132 763331 N/A N/A 47465 47484TCACAATTACATTTTCTTGT 53 2133 763332 N/A N/A 47558 47577TATAGCATGAATTACTTTAC 44 2134 763333 N/A N/A 47651 47670TACCTCCTCTTCAGCAAGGA 78 2135 763334 N/A N/A 47744 47763CCTCTTGTAGTTTTTAAAAT 27 2136 763335 N/A N/A 47951 47970TTGGTTGTTCAAGTGATTCT 33 2137 763336 N/A N/A 48081 48100TCTCACAGTTTTGTTGTTGT 57 2138 763337 N/A N/A 48171 48190CTTTTCATCATGCCTTTATT 40 2139 763338 N/A N/A 48172 48191TCTTTTCATCATGCCTTTAT 40 2140 763339 N/A N/A 48173 48192ATCTTTTCATCATGCCTTTA 37 2141 763340 N/A N/A 48174 48193GATCTTTTCATCATGCCTTT 55 2142 763341 N/A N/A 48175 48194TGATCTTTTCATCATGCCTT 70 2143 763342 N/A N/A 48176 48195TTGATCTTTTCATCATGCCT 51 2144 763343 N/A N/A 48177 48196CTTGATCTTTTCATCATGCC 59 2145 763344 N/A N/A 48178 48197TCTTGATCTTTTCATCATGC 60 2146 763345 N/A N/A 48179 48198CTCTTGATCTTTTCATCATG 38 2147 763346 N/A N/A 48180 48199TCTCTTGATCTTTTCATCAT 43 2148 763347 N/A N/A 48181 48200ATCTCTTGATCTTTTCATCA 36 2149 763348 N/A N/A 48267 48286GGATTACTCCTGGCACAGCT 62 2150 763349 N/A N/A 48360 48379CGATGGAGTACCTACCAACT 36 2151 763350 N/A N/A 48453 48472TACGAGTAGAAGTGACTTGC 53 2152 763351 N/A N/A 48546 48565TCAGTGGAGAGCTATGCAAT 5 2153 763352 N/A N/A 48648 48667TGTAGAATACTTATTTTTTG 28 2154 763353 N/A N/A 48710 48729ATTTTGGATGCTTCTGAAGA 24 2155 763354 N/A N/A 48711 48730TATTTTGGATGCTTCTGAAG 17 2156 763355 N/A N/A 48712 48731GTATTTTGGATGCTTCTGAA 63 2157 763356 N/A N/A 48713 48732TGTATTTTGGATGCTTCTGA 59 2158 763357 N/A N/A 48714 48733TTGTATTTTGGATGCTTCTG 54 2159 763358 N/A N/A 48715 48734TTTGTATTTTGGATGCTTCT 61 2160 763359 N/A N/A 48716 48735GTTTGTATTTTGGATGCTTC 63 2161 763360 N/A N/A 48717 48736GGTTTGTATTTTGGATGCTT 61 2162 763361 N/A N/A 48718 48737TGGTTTGTATTTTGGATGCT 40 2163 763362 N/A N/A 48719 48738ATGGTTTGTATTTTGGATGC 41 2164 763363 N/A N/A 48720 48739GATGGTTTGTATTTTGGATG 55 2165 763364 N/A N/A 48741 48760ACGACATTTTCTTGCCTCTT 74 2166 763365 N/A N/A 48842 48861ATGCTTTCACTTGAAAAAAA 25 2167 763366 N/A N/A 48975 48994CTTTTTTTATTTAAATTCTT 1 2168 763367 N/A N/A 49144 49163ATGGAGAAACTACCCCCATG 27 2169 763368 N/A N/A 49239 49258ACCTCACATGGCAGGAGAAA 32 2170 763369 N/A N/A 49341 49360TTTTTATAAAGAAAGAAGTT 0 2171 763370 N/A N/A 49434 49453TCTTGCTTCTATGTTATATG 65 2172

TABLE 36 Percent reduction of human SNCA mRNA with 5-10-5 MOE gapmerswith mixed internucleoside linkages % SEQ ID SEQ ID SEQ ID SEQ ID Re-SEQ Compound No: 1 No: 1 No: 2 No: 2 duc- ID No start stop start stopSequence (5′ to 3′) tion NO 740410 240 256 4691 4707 GAATTCCTTTACACCAC79 33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC 82 402 762837 238 2574689 4708 TGAATTCCTTTACACCACAC 57 1639 763371 N/A N/A 49527 49546CACCCACCATAAAAGGTGAT 10 2173 763372 N/A N/A 49620 49639TGCCTCTGTTTACAAAGCAA 58 2174 763373 N/A N/A 49713 49732TGGTTTCCTTTGGTGGCTTT 74 2175 763374 N/A N/A 49806 49825TTTCCTTAAGAGGAGCTCTC 53 2176 763375 N/A N/A 49899 49918GATGTAAGTGAGACAGCTCA 56 2177 763376 N/A N/A 49992 50011TTTAGGTAATGGTTTGGTAT 47 2178 763377 N/A N/A 50085 50104ATAGAGGTTATTATTCAGTA 49 2179 763378 N/A N/A 50178 50197AGGAAAACCATCTCTGCTAT 36 2180 763379 N/A N/A 50271 50290GGAAAGAGGTATGAGTGATG 24 2181 763380 N/A N/A 50364 50383GAGTGCTGCCTAAGTCTTGG 50 2182 763381 N/A N/A 50457 50476TTGCTAGCTAAAAGGAGGGT 9 2183 763382 N/A N/A 50550 50569CCAGTTCTAGTTGTACTAGT 52 2184 763383 N/A N/A 50660 50679AAAATGAACTTTTTTATTCG 14 2185 763384 N/A N/A 50753 50772TGCACATCTTTTGCCTGAAA 70 2186 763385 N/A N/A 50846 50865AACTAATCATTATTTTAGAC 0 2187 763386 N/A N/A 50915 50934CATCAATATCTGCAATAATA 63 2188 763387 N/A N/A 50916 50935TCATCAATATCTGCAATAAT 64 2189 763388 N/A N/A 50917 50936TTCATCAATATCTGCAATAA 45 2190 763389 N/A N/A 50918 50937TTTCATCAATATCTGCAATA 64 2191 763390 N/A N/A 50919 50938TTTTCATCAATATCTGCAAT 49 2192 763391 N/A N/A 50920 50939GTTTTCATCAATATCTGCAA 76 2193 763392 N/A N/A 50921 50940GGTTTTCATCAATATCTGCA 60 2194 763393 N/A N/A 50922 50941AGGTTTTCATCAATATCTGC 73 2195 763394 N/A N/A 50923 50942AAGGTTTTCATCAATATCTG 77 2196 763395 N/A N/A 50924 50943AAAGGTTTTCATCAATATCT 65 2197 763396 N/A N/A 50925 50944TAAAGGTTTTCATCAATATC 36 2198 763397 N/A N/A 50926 50945GTAAAGGTTTTCATCAATAT 54 2199 763398 N/A N/A 50939 50958AATTAAGAGGAAGGTAAAGG 2 2200 763399 N/A N/A 51032 51051AAATAATTTCAACATCAGTT 20 2201 763400 N/A N/A 51125 51144CAATAGCTTGCCAAAAATTC 38 2202 763401 N/A N/A 51218 51237ATTTTGTTTCATGGATGTTT 53 2203 763402 N/A N/A 51318 51337AGTCAACATAATTTTTTTTG 34 2204 763403 N/A N/A 51412 51431TCAACAAGGCCTTACTTACG 55 2205 763404 N/A N/A 51505 51524TTATAAAATATCTTCCTAGG 3 2206 763405 N/A N/A 51598 51617TTTTGGCTGCCTCTCAAAAT 21 2207 763406 N/A N/A 51691 51710TTCATTAAAAATTCTGAGTT 3 2208 763407 N/A N/A 51792 51811ATTTTTAATATAATGCTACG 0 2209 763408 N/A N/A 51885 51904CACCAGTGTTTGCATGTCCC 67 2210 763409 N/A N/A 51978 51997CCTCCTACTTCCTAGGCTGC 7 2211 763410 N/A N/A 52071 52090GCTCAATTGGGTGTTCAGCA 62 2212 763411 N/A N/A 52164 52183ACACTGTAAAACTGTCACAA 52 2213 763412 N/A N/A 52310 52329CACATGGATGTATTTGTGCG 49 2214 763413 N/A N/A 52403 52422AGAAGTTTCAAGAACAGTCA 45 2215 763414 N/A N/A 52496 52515TTTAATATACAGATGTTCAG 11 2216 763415 N/A N/A 52589 52608CCCACCTGCCAAAAACACCT 36 2217 763416 N/A N/A 52682 52701TCGAAGTGGGTATGGATGCA 50 2218 763417 N/A N/A 52775 52794GGGCATATGGCTATATACTA 51 2219 763418 N/A N/A 52868 52887TCTAGTTAGCATCTATCCAC 73 2220 763419 N/A N/A 52961 52980TCTTATAAAATTTCTATACT 13 2221 763420 N/A N/A 53054 53073TCATTTTACTTAAGTGGCAC 51 2222 763421 N/A N/A 53147 53166GTCTTTTTCCCATCCTTGAC 53 2223 763422 N/A N/A 53240 53259TTAGCAAGTATAAATATGTT 4 2224 763423 N/A N/A 53333 53352TAGTTGATTGTAGGAAATGT 48 2225 763424 N/A N/A 53426 53445TTGCAAAACAGATGGACTTC 43 2226 763425 N/A N/A 53519 53538TGATGATCTAGCCAAGAGGG 27 2227 763426 N/A N/A 53612 53631ACAAGCTGTACATTAATTAC 42 2228 763427 N/A N/A 53640 53659TTCCATGAAGCCAAGATCAA 46 2229 763428 N/A N/A 53641 53660TTTCCATGAAGCCAAGATCA 62 2230 763429 N/A N/A 53642 53661ATTTCCATGAAGCCAAGATC 63 2231 763430 N/A N/A 53643 53662TATTTCCATGAAGCCAAGAT 66 2232 763431 N/A N/A 53644 53663TTATTTCCATGAAGCCAAGA 61 2233 763432 N/A N/A 53645 53664ATTATTTCCATGAAGCCAAG 53 2234 763433 N/A N/A 53646 53665AATTATTTCCATGAAGCCAA 67 2235 763434 N/A N/A 53647 53666GAATTATTTCCATGAAGCCA 77 2236 763435 N/A N/A 53648 53667TGAATTATTTCCATGAAGCC 66 2237 763436 N/A N/A 53649 53668GTGAATTATTTCCATGAAGC 68 2238 763437 N/A N/A 53650 53669AGTGAATTATTTCCATGAAG 69 2239 763438 N/A N/A 53705 53724AAGTAAGTTCTGAGCTGACA 34 2240 763439 N/A N/A 53798 53817TATTAAGTCTGTTAAGAGGT 54 2241 763440 N/A N/A 53891 53910ATGTTGTATGATGCTCTGGC 74 2242 763441 N/A N/A 53984 54003GTAGATTGCTATTTTGCCAC 55 2243 763442 N/A N/A 54080 54099AATGGGTTTATGTATAATCG 58 2244 763443 N/A N/A 54173 54192CTCCAGACATAGATCTCTCT 63 2245 763444 N/A N/A 54266 54285ACAAGTAAACTGAAACCAGA 23 2246 763445 N/A N/A 54359 54378GTAAGGATGATCATTATAAC 55 2247 763446 N/A N/A 54452 54471ATTAAACATTTTTAATAGCC 27 2248 763447 N/A N/A 54545 54564AGGTGAATAAACTTCGAAAT 51 2249

TABLE 37 Percent reduction of human SNCA mRNA with 5-10-5MOE gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 % ID No start stop startstop Sequence (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 86   33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC93  402 762837 238 257  4689  4708 TGAATTCCTTTACACCACAC 68 1639 763448N/A N/A 54638 54657 TATCAAAAGATTATATATAG  0 2250 763449 N/A N/A 5473154750 TAAATAACATGAATAAGACC 21 2251 763450 N/A N/A 54824 54843TTTTACACATAAGCATATAT 18 2252 763451 N/A N/A 54917 54936AATGAATGTTACCATTTTAT 45 2253 763452 N/A N/A 55011 55030CAATATATTTATTAGGAGAA 30 2254 763453 N/A N/A 55104 55123TCATAAATCAGTCCTCTATA 36 2255 763454 N/A N/A 55197 55216AAAAAGAAGTCAGATATTTC 26 2256 763455 N/A N/A 55290 55309TTTCGGCAGAATTCCAGAGA 56 2257 763456 N/A N/A 55383 55402TGGTTTTCTTTTTCTAGTCA 73 2258 763457 N/A N/A 55476 55495CTCACAAATCATAGGTTTGT 25 2259 763458 N/A N/A 55569 55588GACTATCAATCGGTACTTAT 67 2260 763459 N/A N/A 55663 55682ATTTTATTTGAAATATGTGA 12 2261 763460 N/A N/A 55756 55775GATCTTAGAAATTCATTTAG 42 2262 763461 N/A N/A 55849 55868TTCTCTAAGTACAACACTGC 33 2263 763462 N/A N/A 55942 55961CCACAGTTACATCTGGAAAC 46 2264 763463 N/A N/A 56051 56070AAGTTGTGCGACTTTGGGCA 66 2265 763464 N/A N/A 56144 56163TATCATCAGCAGAACATAGA 36 2266 763465 N/A N/A 56237 56256TAAATATTGTTTTTTCTAAG  1 2267 763466 N/A N/A 56330 56349GACACATTTATATTAGATGT 79 2268 763467 N/A N/A 56423 56442AAGGAGGGAAACAAAGCTCC 22 2269 763468 N/A N/A 56516 56535TACTATATGACATGCTTTCT 31 2270 763469 N/A N/A 56612 56631CAATGGATGAATAGGTGGAT 45 2271 763470 N/A N/A 56705 56724ATGTTGTGGCTTAACCCCAT 54 2272 763471 N/A N/A 56798 56817AAAAAACCTGAAGTACAACA 16 2273 763472 N/A N/A 56891 56910TACTGTGGGTCATTTTTTCT 44 2274 763473 N/A N/A 56987 57006ATAATATCTATATTTAAAAC  0 2275 763474 N/A N/A 57082 57101TATAAAGATGGATTTTTAAA  0 2276 763475 N/A N/A 57175 57194AAATGGATGCTAAGACAATT 35 2277 763476 N/A N/A 57268 57287CCTTCTCTAACTGCCTTTAC 24 2278 763477 N/A N/A 57361 57380GTATAGTTAAAGCTACATTT 59 2279 763478 N/A N/A 57454 57473CAAATTTTGCTTTTACACCC 63 2280 763479 N/A N/A 57547 57566CTTACTTGAGCTAGGTGATC 54 2281 763480 N/A N/A 57640 57659TTCCTCTATTTAATGTATTT 69 2282 763481 N/A N/A 57733 57752TAGCAGTTCCAGGTTCCACA 84 2283 763482 N/A N/A 57826 57845ATCACTTTGGTGTGAGAAGA 14 2284 763483 N/A N/A 57919 57938ATTCCATAGACTTCCAAGTC 60 2285 763484 N/A N/A 58012 58031AGCATCCACATGAAATTGGT 48 2286 763485 N/A N/A 58105 58124GATGTCTTGATACCTTCAGA 79 2287 763486 N/A N/A 58198 58217CTACATGCTAAACTTGTTTT 11 2288 763487 N/A N/A 58291 58310GTGAGAATAAATGTGATCTA 41 2289 763488 N/A N/A 58384 58403CTGTTTCATTAGGAATTTTT 68 2290 763489 N/A N/A 58477 58496TTTATGTACATGGCCAGAAA 32 2291 763490 N/A N/A 58571 58590ACAAAAAATTTCCTAACATT  2 2292 763491 N/A N/A 58664 58683TGTAGCATTTACCTAACAGC 83 2293 763492 N/A N/A 58757 58776AGTGCAGAATCCTGATTGCA 75 2294 763493 N/A N/A 58850 58869CACATTGTAACATAAGCTGT 48 2295 763494 N/A N/A 58943 58962AGTTTGAACTCCGCCCAAGA 32 2296 763495 N/A N/A 59036 59055ACAAGGTTTGCACAAATAAA 48 2297 763496 N/A N/A 59129 59148CCTCATATATAGGGCCTCAC 46 2298 763497 N/A N/A 59222 59241AATTATAAAGCCCTGAAGGC  1 2299 763498 N/A N/A 59315 59334AATGTATTGTTATTTGTCAT 68 2300 763499 N/A N/A 59439 59458CAACTTCTCCATATAACCAA 52 2301 763500 N/A N/A 59592 59611CTAAAGGATGCAAAGGCATA 23 2302 763501 N/A N/A 59685 59704CGTAGATAGAGTTGGAGACC 76 2303 763502 N/A N/A 59788 59807GTGATATATTTACATATATA 60 2304 763503 N/A N/A 59945 59964TTCCAGCGATCCCACTCCTA 26 2305 763504 N/A N/A 60040 60059TTTTTTTACACTGCTGGTAG 31 2306 763505 N/A N/A 60161 60180TTTAATGACCAGGGAAATGC 19 2307 763506 N/A N/A 60418 60437GGGACCTAAAACTATAAAGC 40 2308 763507 N/A N/A 60540 60559CAAAACCTTAAAAATTATAG  0 2309 763508 N/A N/A 60744 60763AAATCCAGAAATAAAGCTAA 18 2310 763509 N/A N/A 60844 60863CTAGATTACCCAACTTCAAA  4 2311 763510 N/A N/A 60972 60991GACTCCCATCAAAATGCCAC 37 2312 763511 N/A N/A 61069 61088ACAGAAACTAATGAAAACAC  0 2313 763512 N/A N/A 61183 61202GAGCCTTTTTACAACAGCTG 61 2314 763513 N/A N/A 61282 61301TTAATTCAGTAAAGTTTCCA 11 2315 763514 N/A N/A 61391 61410AGCATCCAAACTGCTAAAGA 35 2316 763515 N/A N/A 61499 61518TTTCCCCCGAGAACTGGAAT  0 2317 763516 N/A N/A 61592 61611CTGGCATATAAGATACACAC 45 2318 763517 N/A N/A 61691 61710GCAGGAGTAAAAACAAAAAT 29 2319 763518 N/A N/A 61966 61985GTTCCAAAAGATAGAGACAG 37 2320 763519 N/A N/A 62059 62078GTCAGGAAACAAAAAAAGTC 15 2321 763520 N/A N/A 62154 62173CATATATACAAACCTCCTAG 14 2322 763521 N/A N/A 62296 62315AAAGATCTAAACAAGCTCAA 16 2323 763522 N/A N/A 62399 62418CACAAAATACAATACAAAAG  0 2324 763523 N/A N/A 62496 62515AAGATCATACTGTTGCATTC 14 2325 763524 N/A N/A 62674 62693AGGCGGATCACCATAAGTCA  0 2326

TABLE 38 Percent reduction of human SNCA mRNA with 5-10-5MOE gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 % ID No start stop startstop Sequence (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 80   33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC83  402 762837 238 257  4689  4708 TGAATTCCTTTACACCACAC 69 1639 763604N/A N/A 70405 70424 TCATTTCCTTAGTTCCATAT 45 2327 763605 N/A N/A 7049870517 TCTGCCTTGTTTTTTCTTTC 65 2328 763606 N/A N/A 70594 70613TATATAGTTATATATTTACG  0 2329 763607 N/A N/A 70687 70706TTTATATTATATTAACTCTA 13 2330 763608 N/A N/A 70780 70799GGCTTGTCTCTATCCCCTGT 65 2331 763609 N/A N/A 70908 70927GCCCCAGCTTCCGGGTTCAA 24 2332 763610 N/A N/A 71001 71020GAAATATTATTTATACTATT  0 2333 763611 N/A N/A 71094 71113TCTCACAATCACAGAAAACA 31 2334 763612 N/A N/A 71187 71206GACTGCATTTGTATTTCATC 89 2335 763613 N/A N/A 71280 71299AGATTAACAAAATATTAATT 10 2336 763614 N/A N/A 71373 71392TATCATTCTTAACAGAAAAA 20 2337 763615 N/A N/A 71471 71490CTAAGCCATTTTATAACAGG 33 2338 763616 N/A N/A 71568 71587GCCTAACAGGCTATGGACCA 47 2339 763617 N/A N/A 71778 71797CACGGACAGGGATGGTGAGG 52 2340 763618 N/A N/A 71871 71890GTATTGGGCATTATCAGTAA 65 2341 763619 N/A N/A 71964 71983ACTTATCAACACTTAAACTG 23 2342 763620 N/A N/A 72074 72093TAGGTAATTCTAATTTTAAT  6 2343 763621 N/A N/A 72201 72220TACCTAGGTGGTTTCCATAT 46 2344 763622 N/A N/A 72294 72313AATGACAGGGTCTTCTCCTT 58 2345 763623 N/A N/A 72295 72314AAATGACAGGGTCTTCTCCT 66 2346 763624 N/A N/A 72296 72315CAAATGACAGGGTCTTCTCC 60 2347 763625 N/A N/A 72297 72316GCAAATGACAGGGTCTTCTC 76 2348 763626 N/A N/A 72298 72317GGCAAATGACAGGGTCTTCT 68 2349 763627 N/A N/A 72299 72318TGGCAAATGACAGGGTCTTC 83 2350 763628 N/A N/A 72300 72319GTGGCAAATGACAGGGTCTT 77 2351 763629 N/A N/A 72301 72320TGTGGCAAATGACAGGGTCT 89 2352 763630 N/A N/A 72302 72321TTGTGGCAAATGACAGGGTC 73 2353 763631 N/A N/A 72408 72427ACTTTTTCTTTTTAGATTCC 67 2354 763632 N/A N/A 72630 72649TTCTCAACTGCCTGAGTAGC 41 2355 763633 N/A N/A 72756 72775TGTGTGGACTGTGTTTTTTG 70 2356 763634 N/A N/A 72849 72868CAGCTTTTTAGTTCCTCCTA 84 2357 763635 N/A N/A 72942 72961TTCCCCTGTGGCAAGAGCAG 47 2358 763636 N/A N/A 73035 73054ATGCTGTTATAAGATGAATG 58 2359 763637 N/A N/A 73128 73147AAATTATTATAATTCACTCT  5 2360 763638 N/A N/A 73185 73204ACTTTCTGTGTGGTATGTTC 74 2361 763639 N/A N/A 73186 73205GACTTTCTGTGTGGTATGTT 76 2362 763640 N/A N/A 73187 73206AGACTTTCTGTGTGGTATGT 88 2363 763641 N/A N/A 73188 73207CAGACTTTCTGTGTGGTATG 86 2364 763642 N/A N/A 73189 73208ACAGACTTTCTGTGTGGTAT 70 2365 763643 N/A N/A 73190 73209GACAGACTTTCTGTGTGGTA 78 2366 763644 N/A N/A 73191 73210AGACAGACTTTCTGTGTGGT 65 2367 763645 N/A N/A 73192 73211CAGACAGACTTTCTGTGTGG 81 2368 763646 N/A N/A 73193 73212TCAGACAGACTTTCTGTGTG 51 2369 763647 N/A N/A 73194 73213TTCAGACAGACTTTCTGTGT 58 2370 763648 N/A N/A 73195 73214CTTCAGACAGACTTTCTGTG 60 2371 763649 N/A N/A 73221 73240TTGTATTGGTGGAGAAAACA 20 2372 763650 N/A N/A 73314 73333GTGTTGAGAATTTTTCATTG 82 2373 763651 N/A N/A 73407 73426TAGCATCTCTAATGTAGTCT 85 2374 763652 N/A N/A 73500 73519GTAGCTGAATTTCTTCAGCA 22 2375 763653 N/A N/A 73593 73612ATTACAGTGAAATGAAACAT  4 2376 763654 N/A N/A 73686 73705AAATACATTTTGCCTCTGTC 55 2377 763655 N/A N/A 73779 73798CTTTAAGACTTTCCTTAGAC 54 2378 763656 N/A N/A 73872 73891TTTGTTTTAAAACTAGACTT 27 2379 763657 N/A N/A 73965 73984AAAAAGAGATGAAAAGTGTG 22 2380 763658 N/A N/A 74058 74077AGATATGGAGGAGAGTGAAA 28 2381 763659 N/A N/A 74159 74178CTTCCCTCAGCAACAGGCGC 51 2382 763660 N/A N/A 74252 74271GGTCTAGAATCATTCTGAAG 55 2383 763661 N/A N/A 74345 74364AAGGACCTTTCTTCTGAAAG 66 2384 763662 N/A N/A 74438 74457TACACAGAGCACTTCTTATT 41 2385 763663 N/A N/A 74531 74550CTCCCTTTTTCCCACATCTA 48 2386 763664 N/A N/A 74624 74643AAATTAAGTGTTAAGCACAC 60 2387 763665 N/A N/A 74717 74736AAATATTTGCTCAGAGACAC 59 2388 763666 N/A N/A 74810 74829GAATAAAAATGTATAACTAT  6 2389 763667 N/A N/A 75104 75123CAGAGCCTGGCCAAAATGGC 33 2390 763668 N/A N/A 75197 75216AGCACTTAAACAGAAAAAAT 27 2391 763669 N/A N/A 75290 75309TCTATTGTATATTAGGTTGA 67 2392 763670 N/A N/A 75383 75402GATGAAGGAAGAATGATTTT 49 2393 763671 N/A N/A 75476 75495GCTAGTTCATTGTATGTGTC 81 2394 763672 N/A N/A 75569 75588ATTGAATAAAAATTTGTATT  0 2395 763673 N/A N/A 75943 75962CCAGGTATAAAATTTTTTTT 35 2396 763674 N/A N/A 76036 76055GATCTAAGAATACCCCTAGT 25 2397 763675 N/A N/A 76129 76148TTAGATAAAAAGTATACTGT  8 2398 763676 N/A N/A 76222 76241GACAGTTTTCTAATTTTACA 64 2399 763677 N/A N/A 76315 76334GGGTTGGAAATAATACAGAG 43 2400 763678 N/A N/A 76408 76427TTGACCTGCAGTATCTTGAA 28 2401 763679 N/A N/A 76501 76520TACATATTCTTATTCAACTC 46 2402 763680 N/A N/A 76594 76613ATATTATTGATTGTTCTAAA 14 2403

TABLE 39 Percent reduction of human SNCA mRNA with 5-10-5MOE gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SERQ Compound No: 1 No: 1 No: 2 No: 2 % ID No start stop startstop Sequence (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 57   33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC73  402 762837 238 257  4689  4708 TGAATTCCTTTACACCACAC 65 1639 763681N/A N/A 76687 76706 GGATATTCGATTCAAGAACA 56 2404 763682 N/A N/A 7678076799 ATAAATTTGGAAGCTAATGT  2 2405 763683 N/A N/A 76873 76892ACCACATTTTGAAAATAAAG 40 2406 763684 N/A N/A 76966 76985AGCACAGGAAATTAACATAT 62 2407 763685 N/A N/A 77059 77078GTAGGTGTGTTTATTTCTAT 66 2408 763686 N/A N/A 77164 77183CTTTTCATCAGAGATTTTTT 60 2409 763687 N/A N/A 77257 77276GAAATCTAAAAACAGCAAAG  3 2410 763688 N/A N/A 77350 77369GACTATTGTTTTAATGTGTT 48 2411 763689 N/A N/A 77443 77462GGGAGATTTGAGAGAGAGGC 44 2412 763690 N/A N/A 77536 77555ATAGTGGGCTTATGGTGTAC 51 2413 763691 N/A N/A 77630 77649ATTTCTCCATTTCTGTCACT 41 2414 763692 N/A N/A 77738 77757CAGGAGTAAGGACACAGACG 42 2415 763693 N/A N/A 77831 77850CCTCCAGAAAAGGTTTTTAG 62 2416 763694 N/A N/A 77924 77943GAATTGAAACTGCTTAGAAG 28 2417 763695 N/A N/A 78027 78046CCTGACTTTGAATTATTTTG 55 2418 763696 N/A N/A 78120 78139AAAATCAGATAGCAGTGGTG 36 2419 763697 N/A N/A 78213 78232AGGGTACAGAAGGAAAGACA 37 2420 763698 N/A N/A 78306 78325TGAGAGGTGTTTGTTTTGAA 15 2421 763699 N/A N/A 78399 78418TGTTGGCAAGCTTGAAGGGA 44 2422 763700 N/A N/A 78495 78514AATTGAAGGGTTGTAACAGG 29 2423 763701 N/A N/A 78588 78607GCTGGAAAATTAGTCTGTAG 75 2424 763702 N/A N/A 78681 78700CATGGCATGGTCTATACATT 62 2425 763703 N/A N/A 78774 78793AGGTCTCATGGCTGGCAAGT 27 2426 763704 N/A N/A 78867 78886AAACACTTTATCAAATCTTA 41 2427 763705 N/A N/A 78960 78979ATCAGAACAAGTTAAACATT 34 2428 763706 N/A N/A 79053 79072TCTTTTATTCTTGTATCACT 70 2429 763707 N/A N/A 79146 79165TAGCCTTTTGATCTGTTTTT 56 2430 763708 N/A N/A 79239 79258TAAGAATTATGTTAAAACCA 16 2431 763709 N/A N/A 79332 79351CTTAAATTTTAACAATTAAA  0 2432 763710 N/A N/A 79425 79444AATTTACCCCCTAGTAGGCT 57 2433 763711 N/A N/A 79518 79537AGTAACATTTTGAAATGATG 57 2434 763712 N/A N/A 79611 79630CCTGTAGTTCAGTTTTACTG 63 2435 763713 N/A N/A 79704 79723AGATATGAAAATTTTCACTT 25 2436 763714 N/A N/A 79797 79816CTTTTAACTTTAGCTAAATA  0 2437 763715 N/A N/A 79890 79909AGGACCAAAGCTATGGTTAG 52 2438 763716 N/A N/A 79983 80002CAAACAAATAACAGCTTTCA 58 2439 763717 N/A N/A 80076 80095ATAACAAAATTCAGTGCAAC 56 2440 763718 N/A N/A 80169 80188ACATTTAAAGTTTTAACACT 12 2441 763719 N/A N/A 80262 80281GTTTTATAGTTGACAGATGA 53 2442 763720 N/A N/A 80355 80374TCTCTAAATTTGTTGATTTA 26 2443 763721 N/A N/A 80448 80467TGCAGGCACTCACAAACATT 68 2444 763722 N/A N/A 80555 80574ACACCTTTTCTCTTCTTTTT 48 2445 763723 N/A N/A 80648 80667ATCTACTGTTTGAAAGGGTG 61 2446 763724 N/A N/A 80741 80760ACATTGCTCAGAGTTCATGT 44 2447 763725 N/A N/A 80834 80853TAGGTACCATCAGAATTTCA 57 2448 763726 N/A N/A 80927 80946TCATTCTCTGCTACAATAAA 43 2449 763727 N/A N/A 80987 81006GAAATTTTCCAGCTAAAAAA 19 2450 763728 N/A N/A 80988 81007TGAAATTTTCCAGCTAAAAA  0 2451 763729 N/A N/A 80989 81008TTGAAATTTTCCAGCTAAAA 47 2452 763730 N/A N/A 80990 81009CTTGAAATTTTCCAGCTAAA 51 2453 763731 N/A N/A 80991 81010TCTTGAAATTTTCCAGCTAA 47 2454 763732 N/A N/A 80992 81011ATCTTGAAATTTTCCAGCTA 45 2455 763733 N/A N/A 80993 81012AATCTTGAAATTTTCCAGCT 59 2456 763734 N/A N/A 80994 81013AAATCTTGAAATTTTCCAGC 60 2457 763735 N/A N/A 80995 81014TAAATCTTGAAATTTTCCAG 23 2458 763736 N/A N/A 80996 81015ATAAATCTTGAAATTTTCCA 24 2459 763737 N/A N/A 80997 81016CATAAATCTTGAAATTTTCC 40 2460 763738 N/A N/A 81020 81039ATTTCTTTCTCAAGCCCAAA 53 2461 763739 N/A N/A 81113 81132TACATTCCTACTGTATTTAC 38 2462 763740 N/A N/A 81206 81225TGCTTTGATATGGCTTGGAG 64 2463 763741 N/A N/A 81299 81318TGGTATGAGTCACATAAGTA 76 2464 763742 N/A N/A 81392 81411CCTAGAAATTTTGCCTTTTC 40 2465 763743 N/A N/A 81485 81504CTGCAGGTTCTGGAGAGCTG 56 2466 763744 N/A N/A 81578 81597TGTTTACTGCCACTATTCAC 53 2467 763745 N/A N/A 81681 81700CTAACTGAACTTTTAAAAAT  4 2468 763746 N/A N/A 81774 81793AATACAATCTATCAGCATTA 53 2469 763747 N/A N/A 81868 81887AATTTTGGAGGAATTTATTT  0 2470 763748 N/A N/A 81961 81980TTGTGCTTCAATAATACCAA 37 2471 763749 N/A N/A 82112 82131TGGGTTTCATGGTGTTAGCT 69 2472 763750 N/A N/A 82237 82256GTAGGCTCAGTGCAAACTCT 57 2473 763751 N/A N/A 82330 82349AGTCTTTTTACATTATAATA 28 2474 763752 N/A N/A 82423 82442TAACAGATTTGTGGTGAAAA 52 2475 763753 N/A N/A 82516 82535AACCATAAGAGAGGACAAAC 39 2476 763754 N/A N/A 82609 82628AATGATCTTTAAAACATTCA  9 2477 763755 N/A N/A 82702 82721GAGGACAATAAAATGACCTT 70 2478 763756 N/A N/A 82810 82829CTCCTCTCAACTGCCAGCGC 52 2479 763757 N/A N/A 82903 82922TTTACTAAGTCATCTGTGAA 19 2480

TABLE 40 Percent reduction of human SNCA mRNA with 5-10-5MOE gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 % ID No start stop startstop Sequence (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 75   33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC84  402 762837 238 257  4689  4708 TGAATTCCTTTACACCACAC 63 1639 763758N/A N/A 82996 83015 CTTATGAGCTGTTTAGGAAG 45 2481 763759 N/A N/A 8308983108 TGTCAACTCTGCCAATGTGA 56 2482 763760 N/A N/A 83183 83202AATAAATGCTATGTAATTTA  3 2483 763761 N/A N/A 83276 83295GAAGGGTTGCTATGATAGTT 56 2484 763762 N/A N/A 83369 83388TGAATTCTAACCAAAAGCTT 33 2485 763763 N/A N/A 83462 83481GACAAATGTGTCACTTTTTA 11 2486 763764 N/A N/A 83555 83574AAATTCATGAGGAATGCAAT 41 2487 763765 N/A N/A 83648 83667CATACAATATTTTTGACAGA 50 2488 763766 N/A N/A 83741 83760GTGACGCACATTTACACCAG 61 2489 763767 N/A N/A 83834 83853AAAGATTTTTATCTTAGCCT 42 2490 763768 N/A N/A 83927 83946CCATTTACAAAGATGACCAG 22 2491 763769 N/A N/A 84020 84039TCAAAGTAGTGAATTACATC 51 2492 763770 N/A N/A 84113 84132TGTTTGGACTTATAAACTAT 55 2493 763771 N/A N/A 84206 84225TAATGGGCAAGCAAAAAATT  0 2494 763772 N/A N/A 84552 84571TCATGTTGCCTAGGCTAGAA 58 2495 763773 N/A N/A 84645 84664CTAGATAACATACAATATAA  0 2496 763774 N/A N/A 84752 84771GAGAAATTATTATATTTTAT  0 2497 763775 N/A N/A 84845 84864AGACTACAAAATTGCTAAAA 32 2498 763776 N/A N/A 84938 84957CAAAAAGATTTTTATGGAGT  3 2499 763777 N/A N/A 85031 85050AATTTAAGTTTAAATATTCT  0 2500 763778 N/A N/A 85124 85143CCTCATTTTGCCAGTATTAA 76 2501 763779 N/A N/A 85217 85236ATGGGAAAATTGTGACTGTT 59 2502 763780 N/A N/A 85315 85334GCAAATATATGAATTTTTTA 12 2503 763781 N/A N/A 85424 85443TCATTGGAAAATCTTGAACG 61 2504 763782 N/A N/A 85517 85536ATTTGTAGACCTATGTTGAA 10 2505 763783 N/A N/A 85610 85629GAATAATAAGATTCAGTCAT 56 2506 763784 N/A N/A 85703 85722GAAACCATTAAAATATTTAT  0 2507 763785 N/A N/A 85797 85816TCTAAGTTTTTATTAATTAA  0 2508 763786 N/A N/A 85891 85910ATGATTAGGATTTTTATTTC  1 2509 763787 N/A N/A 85984 86003TTTATATTTAAATCACACAA  0 2510 763788 N/A N/A 86077 86096AATTGCTGTTTTAATCATGA 54 2511 763789 N/A N/A 86170 86189CAGATTTATCTACTTGAAAC 46 2512 763790 N/A N/A 86263 86282GTAGAGTTTTTGGTCAGTGG 63 2513 763791 N/A N/A 86356 86375TTTTTGTTCTTGGGATGTTG 53 2514 763792 N/A N/A 86449 86468TAACTTTCAACCGTGAAAAA 15 2515 763793 N/A N/A 86542 86561GTCTGTTTTCTAACTAGCTT 76 2516 763794 N/A N/A 86635 86654GCACATTGTCAAATAAACAA 61 2517 763795 N/A N/A 86728 86747CAGGAATTATCCAAAGTCAC 70 2518 763796 N/A N/A 86821 86840ACCTGGGTTAAGTAAATGGC 38 2519 763797 N/A N/A 86914 86933CACTGGAGAGACTGTGAAGG 53 2520 763798 N/A N/A 87007 87026AGCAGCAGATTTCAAAAGGG 69 2521 763799 N/A N/A 87100 87119TTTTGATTGTGGTAATTGGA 36 2522 763800 N/A N/A 87193 87212TACAAGAGTGGAAATGGCTG 27 2523 763801 N/A N/A 87286 87305CCGTTACATGCTCTCTAATT 46 2524 763802 N/A N/A 87379 87398CTCCTGATCTCAATTGAAAT 10 2525 763803 N/A N/A 87472 87491AAGCCTTCATATACGAGTTT 60 2526 763804 N/A N/A 87565 87584TATCTCCAGCCTTCACCTCT 13 2527 763805 N/A N/A 87658 87677TCTCCTTCTTACAAAATCCA 53 2528 763806 N/A N/A 87759 87778GTTGTTCTTCTTCTTATTAT 58 2529 763807 N/A N/A 87854 87873GTTAAAATTTGAAATAATGA  0 2530 763808 N/A N/A 87940 87959AGTTGTGACCATGCAATAAA 50 2531 763809 N/A N/A 87941 87960AAGTTGTGACCATGCAATAA 51 2532 763810 N/A N/A 87942 87961TAAGTTGTGACCATGCAATA 44 2533 763811 N/A N/A 87943 87962GTAAGTTGTGACCATGCAAT 65 2534 763812 N/A N/A 87944 87963AGTAAGTTGTGACCATGCAA 69 2535 763813 N/A N/A 87945 87964TAGTAAGTTGTGACCATGCA 68 2536 763814 N/A N/A 87946 87965TTAGTAAGTTGTGACCATGC 55 2537 763815 N/A N/A 87947 87966ATTAGTAAGTTGTGACCATG 44 2538 763816 N/A N/A 87948 87967CATTAGTAAGTTGTGACCAT 42 2539 763817 N/A N/A 87949 87968CCATTAGTAAGTTGTGACCA 71 2540 763818 N/A N/A 87950 87969CCCATTAGTAAGTTGTGACC 71 2541 763819 N/A N/A 88040 88059GCTATCAAGACATTATGTAG 58 2542 763820 N/A N/A 88133 88152ATCTATGAAAGCAAATGTTT 35 2543 763821 N/A N/A 88227 88246CTTTTTTAAACAAAATACAG  0 2544 763822 N/A N/A 88320 88339ATGCTACAAGCAGGCACTTA 22 2545 763823 N/A N/A 88413 88432TCTGTTATCTTAAGAGGCTT 76 2546 763824 N/A N/A 88506 88525TGGACTTTATTGCTCAAAGC 65 2547 763825 N/A N/A 88599 88618CAGACAAAAACATCCGATAT 28 2548 763826 N/A N/A 88692 88711CCCTAGACAACTATCACCTG  9 2549 763827 N/A N/A 88785 88804TGGAAGCCCTGAGGAAGTGG 59 2550 763828 N/A N/A 88878 88897AACAGCAAGGACAATGTCTA 53 2551 763829 N/A N/A 88971 88990GCCATGTGTTATATACTTTG 73 2552 763830 N/A N/A 89075 89094ATTAGGTAGATTTTTTTTAA  0 2553 763831 N/A N/A 89169 89188ATGATGGTGAATAAATTAAA 11 2554 763832 N/A N/A 89262 89281AGAAAATGCTTTAAGCTCAT 57 2555 763833 N/A N/A 89355 89374AAAAGATAAATTGCTAGGTT 16 2556 763834 N/A N/A 89452 89471ACTAATTAATTAGTTGAATA  8 2557

TABLE 41 Percent reduction of human SNCA mRNA with 5-10-5MOE gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 % ID No start stop startstop Sequence (5′ to 3′) Reduction NO 740410 240 256  4691  4707GAATTCCTTTACACCAC 67   33 741410 N/A N/A 87946 87962 GTAAGTTGTGACCATGC83  402 762837 238 257  4689  4708 TGAATTCCTTTACACCACAC 76 1639 763835N/A N/A 89545 89564 TCAATTCTCTTTAGAATTTC 27 2558 763836 N/A N/A 8963889657 AGAGATTCATGCTCATTTAT 45 2559 763837 N/A N/A 89731 89750ATCTATCCACTCTCCTATAG  0 2560 763838 N/A N/A 89824 89843ACTTTTTTATTAGAGCCCCC 14 2561 763839 N/A N/A 89917 89936GACTACATGTCCTTTAAATG 64 2562 763840 N/A N/A 90015 90034AAGACTGCAGGCTTGAGCCA 23 2563 763841 N/A N/A 90141 90160ATGCCACCACATCCCACTTT 21 2564 763842 N/A N/A 90312 90331TGTTATATTTTAAAAGTTTC  0 2565 763843 N/A N/A 90439 90458ATATACACAAAAGCAGATAT 19 2566 N/A N/A 90405 90424 763844 N/A N/A 9050090519 AAATATATGTGTAAATACAC  0 2567 763845 N/A N/A 90593 90612CTCCCATTCTCTCTCTCTAC 35 2568 763846 N/A N/A 90686 90705CCCAGAAACTAACATCTTCT 48 2569 763847 N/A N/A 90779 90798CAGGAAAAAGAATACTTTCT 34 2570 763848 N/A N/A 90945 90964TCCTGCCACCACACCCACTA 11 2571 763849 N/A N/A 91047 91066TAGCTATAGTGCAATGGCGC  7 2572 763850 N/A N/A 91140 91159TTTCATAACTGTATGATTTG 33 2573 763851 N/A N/A 91233 91252ACCATTAAAAGTTTAGTGGA 43 2574 763852 N/A N/A 91326 91345ATGTGCATGCCAGTGTGTTA 34 2575 763853 N/A N/A 91419 91438GATAAAGAAGGAATGCACAA 34 2576 763854 N/A N/A 91520 91539CTTACTTTCTTGCAAAAGGG 43 2577 763855 N/A N/A 91614 91633GAAAATAAAAAGGCAGCTTT 13 2578 763856 N/A N/A 91707 91726TAATAGTGAATGTTGTTTTA 21 2579 763857 N/A N/A 91800 91819CATGCATCTAAAGATAACTG 33 2580 763858 N/A N/A 91893 91912TCCTAGGCTTTGTCTCTTAA 38 2581 763859 N/A N/A 91986 92005TTTAAAACTTTATCTTCCTT 35 2582 763860 N/A N/A 92079 92098AGATACTGTTGCCCCAAGTA 48 2583 763861 N/A N/A 92172 92191TACTAAAAAAAACCACTAAC  0 2584 763862 N/A N/A 92265 92284CCACTGTCTAACAAATAATG 32 2585 763863 N/A N/A 92358 92377ATGATTGGTGTAAGCGAATG 24 2586 763864 N/A N/A 92451 92470ATTATCCTTCAACAGAGCTA 21 2587 763865 N/A N/A 92544 92563GCCCATCCTTAGATCTTAGT 46 2588 763866 N/A N/A 92642 92661CGAGTGACTCAGTTTCCTTA 64 2589 763867 N/A N/A 92735 92754CCTTCACTTTGGAGGATGCG 37 2590 763868 N/A N/A 92828 92847CCTAGAGGGTGCCTTCCCAG 28 2591 763869 N/A N/A 92921 92940ATATTTACACTGCTTCATAA  3 2592 763870 N/A N/A 93014 93033TTATGACCTGTAATGTACTT 25 2593 763871 N/A N/A 93151 93170CAAAAGACAAGCACACACAC  0 2594 763872 N/A N/A 93244 93263TAAGTATTTTTAGTACTTTA 14 2595 763873 N/A N/A 93337 93356GAGGGACTTTTGCAATTGTC 15 2596 763874 N/A N/A 93430 93449GAATCAAAATAAGAGGTCAA 35 2597 763875 N/A N/A 93521 93540TTTTGAGTTCCAGGGATTCA 54 2598 763876 N/A N/A 93522 93541GTTTTGAGTTCCAGGGATTC 70 2599 763877 N/A N/A 93523 93542TGTTTTGAGTTCCAGGGATT 74 2600 763878 N/A N/A 93524 93543ATGTTTTGAGTTCCAGGGAT 50 2601 763879 N/A N/A 93525 93544AATGTTTTGAGTTCCAGGGA 57 2602 763880 N/A N/A 93526 93545CAATGTTTTGAGTTCCAGGG 57 2603 763881 N/A N/A 93527 93546GCAATGTTTTGAGTTCCAGG 61 2604 763882 N/A N/A 93528 93547AGCAATGTTTTGAGTTCCAG 68 2605 763883 N/A N/A 93529 93548CAGCAATGTTTTGAGTTCCA 70 2606 763884 N/A N/A 93530 93549TCAGCAATGTTTTGAGTTCC 66 2607 763885 N/A N/A 93531 93550TTCAGCAATGTTTTGAGTTC 33 2608 763886 N/A N/A 93621 93640GCATTTCTTAATTTTTTTAT  6 2609 763887 N/A N/A 93714 93733TTGTCTGCTACTATTTTTTC 25 2610 763888 N/A N/A 93807 93826TTTAATATTTATGAATGTGA 15 2611 763889 N/A N/A 93900 93919AAGCCTTTATTTTTTATTGC  8 2612 763890 N/A N/A 93993 94012ATGAGGGCAAGCTGGCTTTT 40 2613 763891 N/A N/A 94086 94105CAAGGAGATTGAGTTTACCA 51 2614 763892 N/A N/A 94179 94198CAAAGCATTCTTGCTTGCTC 50 2615 763893 N/A N/A 94272 94291TAATTTATGTCAGTCATTAA  0 2616 763894 N/A N/A 94365 94384GGCTGCCGAAAGCAGGAAAA 32 2617 763895 N/A N/A 94458 94477TTTAAATGTCACAGCTATTT 15 2618 763896 N/A N/A 94551 94570CTTCCCCTAAATCTCTCTGT 20 2619 763897 N/A N/A 94644 94663AGTTAACAAATTAATGAAAC 10 2620 763898 N/A N/A 94993 95012AACTTCAGTTTTGTGGCGGG 15 2621 763899 N/A N/A 95086 95105CCTGGAAAATGAGGACTTTC 44 2622 763900 N/A N/A 95179 95198ACTGATTAAGAAATGTGAGG 31 2623 763901 N/A N/A 95272 95291TGAAAGCCACCGTGATGAAC  4 2624 763902 N/A N/A 95365 95384AGATTAAAGCGATTCCTGCT 12 2625 763903 N/A N/A 95459 95478CTTAGTATCATCATCATCAC 39 2626 763904 N/A N/A 95552 95571CCCAGAAAATAAGCAGACTG 45 2627 763905 N/A N/A 95645 95664GCAAATACAATATTTGAAAG  0 2628 763906 N/A N/A 95738 95757GATCAGAATGACCAGTGCAC 43 2629 763907 N/A N/A 95831 95850TCAAACTATAATTTGGTGTC 61 2630 763908 N/A N/A 95924 95943TCTAGAGAATGATTCATCTT 39 2631 763909 N/A N/A 96017 96036TACCCTCTTGCTATACAAAC 30 2632 763910 N/A N/A 96110 96129GATGAAAATTGAAATTTGAT 13 2633 763911 N/A N/A 96203 96222TTAAAAATAACTGTATTTGG  0 2634

TABLE 42 Percent reduction of human SNCA mRNA with 5-10-5MOE gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 % ID No start stop startstop Sequence (5′ to 3′) Reduction NO 740410 240 256   4691   4707GAATTCCTTTACACCAC 74   33 741410 N/A N/A  87946  87962 GTAAGTTGTGACCATGC80  402 762837 238 257   4689   4708 TGAATTCCTTTACACCACAC 67 1639 763912N/A N/A  96296  96315 ATTAACTAAAAACCAAGTCT  7 2635 763913 N/A N/A  96391 96410 CAAAGCTGCAGACCATTTTG 25 2636 763914 N/A N/A  96484  96503ACAGGCAAAGAATTTTTGAG 35 2637 763915 N/A N/A  96577  96596AAACAACTGCTGAGAAGCAG  0 2638 763916 N/A N/A  96670  96689TTTAATTGCTGTTGTGTGGT 45 2639 763917 N/A N/A  96763  96782TAAGACATAAATGTCAGAGG 25 2640 763918 N/A N/A  96858  96877TGAAGATGAAGAAAGGAAAG  2 2641 763919 N/A N/A  96951  96970GAGCCAATAACAGAGATGAT 36 2642 763920 N/A N/A  97044  97063AGACTGTTAATGTAGTAGGA 29 2643 763921 N/A N/A  97137  97156GGACAATTAATTTTGAGGGT 48 2644 763922 N/A N/A  97230  97249CTTCAGGAGATAAAGGAACC  7 2645 763923 N/A N/A  97323  97342TTATGCTTCAGGGATGCATA 36 2646 763924 N/A N/A  97416  97435TTTACTAAGTAATTGGTACT 26 2647 763925 N/A N/A  97509  97528AAGGCAGCAAAGAGGTAAAA  2 2648 763926 N/A N/A  97602  97621GGTAAGTCATCAGAGTTCAT 27 2649 763927 N/A N/A  97695  97714TTGAGTCTGAGATGCCTCCA 34 2650 763928 N/A N/A  97788  97807TTTGAGCTTGACCAACTAGG 45 2651 763929 N/A N/A  97881  97900GCAGTTACTGACTTGCTTGA 38 2652 763930 N/A N/A  97974  97993GCTGCAAGCACACCTGCCTT 36 2653 763931 N/A N/A  98067  98086AAGAGGAACGCAGAGCTCAG  8 2654 763932 N/A N/A  98160  98179GAGTATCATGATTTTCTTGC 57 2655 763933 N/A N/A  98253  98272CAAGCCTGCCAGTCTTTTGA 43 2656 763934 N/A N/A  98346  98365TATAGGTGCAAACTACAAGT 35 2657 763935 N/A N/A  98439  98458GGAATACAGCCAAAAACTTG 13 2658 763936 N/A N/A  98532  98551AGCTACATTCAAGTCTGCAA 57 2659 763937 N/A N/A  98803  98822CGAATGGGCGGATCACAAGG  7 2660 763938 N/A N/A  98896  98915AAGAATCGAAACTAAAAACC  9 2661 763939 N/A N/A  98989  99008AATGTATATCATATATTGTC 57 2662 763940 N/A N/A  99082  99101GACCCATGCACAGTCATAAT 36 2663 763941 N/A N/A  99175  99194CGTAAATGTTTCAACTGAAA 45 2664 763942 N/A N/A  99268  99287GTTGGAAGCTCAGGAGAAAA 56 2665 763943 N/A N/A  99361  99380TTGTTGAGGAACTGAAATTG 20 2666 763944 N/A N/A  99454  99473AGTGGGCTTGTGGTATTTGT  7 2667 763945 N/A N/A  99547  99566GCAAAGGGAGAACAAACAAA  0 2668 763946 N/A N/A  99641  99660CTGTATATAATTTTTTCAAC 35 2669 763947 N/A N/A  99734  99753ACTAAATGTTTATTTGCATT 44 2670 763948 N/A N/A  99827  99846GAATTTAAAGAGGAATAAAA  0 2671 763949 N/A N/A  99920  99939AATTTCATTATGATTATCGC 64 2672 763950 N/A N/A 100013 100032TCTTCAAACCTTTTGACCAA 41 2673 763951 N/A N/A 100111 100130GAAATAAATTGTTCATTTTG  7 2674 763952 N/A N/A 100205 100224GAAAAAATAGTTTATTATAA  0 2675 763953 N/A N/A 100298 100317TATATGATTTTTTGCAAGGG 41 2676 763954 N/A N/A 100394 100413GTTAAAGGAAATGTTTATAT 12 2677 763955 N/A N/A 100487 100506AAATTAATCCTTTCCAAATG  0 2678 763956 N/A N/A 100580 100599AATATTAGTTGTCAAATGTC 42 2679 763957 N/A N/A 100673 100692CTCTTTGAGGAAGTTACTAC 23 2680 763958 N/A N/A 100766 100785AATAACAATAACAGTTAATG  0 2681 763959 N/A N/A 100860 100879GATTATCAAGAAAGATAATG  0 2682 763960 N/A N/A 100953 100972GCTACTTTCTTTCAGTTACC 49 2683 763961 N/A N/A 101046 101065GCCAGAGGACCATAGTGGTT 45 2684 763962 N/A N/A 101143 101162TACTAAGTGAAGTTTGAGGG 18 2685 763963 N/A N/A 101236 101255AGAAAGGCTTTAAGATAGCT  9 2686 763964 N/A N/A 101329 101348AAGGATGGGCTCTGAAGCAG 13 2687 763965 N/A N/A 101422 101441CCCAGGAGTTTGCTCTCAAA 36 2688 763966 N/A N/A 101515 101534AGAGTCTGCTTTCATATTTT 36 2689 763967 N/A N/A 101914 101933TGGAGGCAGGTCTTTTTTTT 32 2690 763968 N/A N/A 102007 102026ACGATGTGAAGATGGGTCAA 45 2691 763969 N/A N/A 102100 102119TTAAACTATATTCAAATTTG  0 2692 763970 N/A N/A 102193 102212AATGCACAAAGGGAAATCTG 38 2693 763971 N/A N/A 102286 102305AATTAGCTGACTCACCTAAT  4 2694 763972 N/A N/A 102379 102398AGCAAAGAGGTAGTATGCTG 61 2695 763973 N/A N/A 102472 102491GTTTAAATACATTCAACCAT 46 2696 763974 N/A N/A 102565 102584GGTTTGGCAGTGGAGGAGAG 28 2697 763975 N/A N/A 102658 102677CCCTTCTAGCTGTTTCTTTA 40 2698 763976 N/A N/A 102831 102850TATAGAGATGAAGTTTCATT 29 2699 763977 N/A N/A 102982 103001CCCTATTGCCCAGGCTGTAA 21 2700 763978 N/A N/A 103075 103094CTTTAGAGAACCCAGTCTTA 38 2701 763979 N/A N/A 103175 103194ATAGTCACATTGGTGAACGC 33 2702 763980 N/A N/A 103268 103287TTGCTCTCCCTCAGTTATGT 52 2703 763981 N/A N/A 103361 103380TGCTATTATATATGCTAAGC 54 2704 763982 N/A N/A 103454 103473CTGATGATCTCTGGTGCCAC 35 2705 763983 N/A N/A 103547 103566CTACTAACCTGTAAAAGACA  1 2706 763984 N/A N/A 103640 103659ACTTTACAACAAGATAAAAA  0 2707 763985 N/A N/A 103733 103752TCTGGTACAGTCCTACTACC 61 2708 763986 N/A N/A 103826 103845AATATAATTTATAGCATTAC  0 2709 763987 N/A N/A 103919 103938TGAGGCAATATGCAGACGAA 51 2710 763988 N/A N/A 104012 104031TTTAGAAATGCATCAAAGTG 18 2711

TABLE 43 Percent reduction of human SNCA mRNA with 5-10-5MOE gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 1 No: 1 No: 2 No: 2 % ID No start stop startstop Sequence (5′ to 3′) Reduction NO 740410 240 256   4691   4707GAATTCCTTTACACCAC 89   33 741410 N/A N/A  87946  87962 GTAAGTTGTGACCATGC90  402 762837 238 257   4689   4708 TGAATTCCTTTACACCACAC 79 1639 763989N/A N/A 104105 104124 CAAACTTTAATTTTTGGGAA 31 2712 763990 N/A N/A 104198104217 TGTATCCAATGCCCAAAGGA 49 2713 763991 N/A N/A 104291 104310CATTTATTGTTTACATACTC 29 2714 763992 N/A N/A 104384 104403GCAAAACAATATGCATATTT 63 2715 763993 N/A N/A 104477 104496GAGGTCTTGTTTTGGAAAGG 54 2716 763994 N/A N/A 104570 104589TGGATACTCTGATTTCTCTT 76 2717 763995 N/A N/A 104663 104682AGCAAAGGGCATCTGATTCA 46 2718 763996 N/A N/A 104756 104775ACCTTGTTAAAAAGCAAGGT  2 2719 763997 N/A N/A 104849 104868GGAGTGTGTACATAGTGTAG 46 2720 763998 N/A N/A 104942 104961AAAATGAAATCAAGCCCAGA 23 2721 763999 N/A N/A 105035 105054GAGATAGTAGCCAAAAAGAT 22 2722 764000 N/A N/A 105128 105147TTGTTTTGCTGCATTATTGA 42 2723 764001 N/A N/A 105221 105240CAACTTTCACAGCCTTAAAC 38 2724 764002 N/A N/A 105314 105333TTTGGAGCAATGTGATGTTT 40 2725 764003 N/A N/A 105407 105426GAGCTGCAGCAAGTTTTTTC 56 2726 764004 N/A N/A 105502 105521GCTGCTCTTTGAGAAAGTTC 64 2727 764005 N/A N/A 105595 105614AAGAAAAATTGAAATTCAAG  9 2728 764006 N/A N/A 105688 105707AAAATAGCAAGGTTTCATCA 17 2729 764007 N/A N/A 105781 105800TTAAAAAAGATATGCTCATT 12 2730 764008 N/A N/A 105874 105893CACTGCCCGACATCACCAAT 20 2731 764009 N/A N/A 105967 105986AACCACACTCTTCTAGAATC 41 2732 764010 N/A N/A 106060 106079TAAGGAAATTATCTTTATTC 27 2733 764011 N/A N/A 106153 106172TGTCTTTAGGAATACAACTA 55 2734 764012 N/A N/A 106246 106265GCTAATAGCTTATTGGGAAG 47 2735 764013 N/A N/A 106339 106358GGGTTGAATAGCTGATATAA 65 2736 764014 N/A N/A 106432 106451AGACTTAAAAGCTATATTAG  6 2737 764015 N/A N/A 106525 106544TCAGTTCAGTATCTTATATC 68 2738 764016 N/A N/A 106618 106637ACCTTTTATTCTCTCTCTAC 60 2739 764017 N/A N/A 106713 106732TAAAATAAATGAGAAAAACG  2 2740 764018 N/A N/A 106806 106825CCAATATAACAAATGTTAAA 24 2741 764019 N/A N/A 106899 106918GAGTATTCATGACTTGTTTT 53 2742 764020 N/A N/A 106999 107018TGTTATCTATAAAGAAATAT 17 2743 764021 N/A N/A 107092 107111AATATAACAACAAACACTTC  2 2744 764022 N/A N/A 107185 107204TATGTTTTTCTGAATATGTG 36 2745 764023 N/A N/A 107278 107297GCAATTTCAGGTGTCCTAGT 79 2746 764024 N/A N/A 107434 107453CCCACATAACTTTTATTACA 42 2747 764025 N/A N/A 107539 107558CGTGGTTTTGTTTTCCATGG 70 2748 764026 N/A N/A 107641 107660ATCTATCTAGGTTTGGGTGG 57 2749 764027 N/A N/A 107734 107753TTATTTCTTTAGGTGTGATG 49 2750 764028 N/A N/A 107827 107846ATTCTCATTGGGAACCCTAC 40 2751 764029 N/A N/A 107920 107939GTAAATTGCAACTAAAAAGA  8 2752 764030 N/A N/A 108013 108032AAACATGTTCATTGCTTACA 57 2753 764031 N/A N/A 108271 108290GGTTCTCCTATAGTCCCAGC 58 2754 764032 N/A N/A 108364 108383ACATGATCGGTGAGGTCAGG 40 2755 764033 N/A N/A 108457 108476TCGACAATAGGGTTTACGAC 56 2756 764034 N/A N/A 108550 108569TTAAGTGGGCTATTGTTCAC 39 2757 764035 N/A N/A 108643 108662ACTATTGATGAAGTTAAGTG 14 2758 764036 N/A N/A 108736 108755TGCCATAGGACTTAATTCTT 69 2759 764037 N/A N/A 108857 108876TTGACTTGTTTGTATTAATC 62 2760 764038 N/A N/A 108970 108989CCTGCAGTAATGGAACAGCG 67 2761 764039 N/A N/A 109063 109082TGAACTTTGAAGGATGTACA 40 2762 764040 N/A N/A 109156 109175CTACCCTGTTTGTTGTTTGA 15 2763 764041 N/A N/A 109249 109268TTTTCCATGATTTTGAAACT 30 2764 764042 N/A N/A 109342 109361ACAACAGGGAGAAGGAAACG 17 2765 764043 N/A N/A 109435 109454ATGACAGAGCTTTTGTGATG 35 2766 764044 N/A N/A 109528 109547TTCACTTCTTGGTAGATACG 45 2767 764045 N/A N/A 109627 109646CTAAAAAAAATCCAAATAAT 14 2768 764046 N/A N/A 109720 109739AGTAAGAAAAGGTCACACTA 34 2769 764047 N/A N/A 109813 109832ATTTTCAACAACATGTCTGA 14 2770 764048 N/A N/A 109906 109925AAAGGTAAGTGAAAATTCAA 29 2771 764049 N/A N/A 109999 110018CTCAGCCTGAAATGGTCATG 39 2772 764050 N/A N/A 110092 110111AGACTGAGACTATACATATT 29 2773 764051 N/A N/A 110185 110204AACTTTTATAACCACTTATA 31 2774 764052 N/A N/A 110278 110297AAAGGTAAAAAGTTTGGAAG  6 2775 764053 N/A N/A 110371 110390ATTATGTAACAACTACCTAT  8 2776 764054 N/A N/A 110624 110643GTTACAATGAAACCCCATCT 29 2777 764055 N/A N/A 110724 110743TTATTGCTGGGTGCAGTGGT 24 2778 764056 N/A N/A 110817 110836CCAAAGATATTTTTCACAAG 65 2779 764057 N/A N/A 110910 110929AAACATTGCGGCAACATGGG 22 2780 764058 N/A N/A 111003 111022AAATCTTACATATAGGGATG 39 2781 764059 N/A N/A 111097 111116TCCTTCTTCATTCTAATATT  7 2782

TABLE 44 Percent reduction of human SNCA mRNA with 5-10-5MOE gapmers with mixed internucleoside linkages SEQ ID SEQ ID SEQ IDSEQ ID SEQ Compound No: 2 No: 2 No: 5 No: 5 % ID No start stop startstop Sequence (5′ to 3′) Reduction NO 740410  4691  4707 N/A N/AGAATTCCTTTACACCAC 89   33 741410 87946 87962 N/A N/A GTAAGTTGTGACCATGC90  402 762837  4689  4708 N/A N/A TGAATTCCTTTACACCACAC 79 1639 764060N/A N/A 38 57 CTTTACACCACACTCTTTCA 23 2783 764061 N/A N/A 39 58CCTTTACACCACACTCTTTC 45 2784 764062 N/A N/A 40 59 TCCTTTACACCACACTCTTT56 2785 764063 N/A N/A 41 60 TTCCTTTACACCACACTCTT 52 2786 764064 N/A N/A42 61 ATTCCTTTACACCACACTCT 70 2787 764065 N/A N/A 43 62AATTCCTTTACACCACACTC 67 2788

Example 6: Design of Gapmers with Mixed Internucleoside LinkagesComplementary to Human SNCA

Modified oligonucleotides complementary to a human SNCA nucleic acidwere designed. The modified oligonucleotides in Table 45 are gapmers.The gapmers have a central gap segment that comprises2′-deoxynucleosides and is flanked by wing segments on both the 5′ endon the 3′ end comprising 2′-MOE nucleosides and cEt nucleosides. Allcytosine residues throughout each gapmer are 5-methyl cytosines. Theinternucleoside linkages are mixed phosphodiester internucleosidelinkages and phosphorothioate internucleoside linkages. The sequence andchemical notation column specifies the sequence, including 5′-methycytosines, sugar chemistry, and the internucleoside linkage chemistry,wherein subscript ‘d’ represents a 2′-deoxyribose sugar; subscript ‘e’represents a 2′-MOE modified sugar; subscript ‘k’ represents a cEtmodified sugar; subscript ‘o’ represents a phosphodiesterinternucleoside linkage; subscript ‘s’ represents a phosphorothioateinternucleoside linkage; and a ‘m’ superscript before the cytosineresidue indicates a 5-methyl cytosine. “Start Site” indicates the5′-most nucleoside to which the gapmer is complementary in the humannucleic acid sequence. “Stop Site” indicates the 3′-most nucleoside towhich the gapmer is complementary in the human nucleic acid sequence.

Each modified oligonucleotide listed in the Tables below iscomplementary to human SNCA nucleic acid sequences SEQ ID NO: 2 or SEQID NO:5, as indicated. ‘N/A’ indicates that the modified oligonucleotideis not complementary to that particular nucleic acid with 100%complementarity.

TABLE 45 Modified oligonucleotides complementary to human SNCA mRNASEQ ID SEQ ID SEQ ID SEQ ID SEQ Compound No: 2 No: 2 No: 5 No: 5Sequence and chemical notation ID No start stop start stop (5′ to 3′)NO: 788813 N/A N/A  4686  4702 ^(m)C_(es)^(m)C_(eo)T_(eo)T_(eo)T_(ds)A_(ds) ^(m)C_(ds)A_(ds) ^(m)C_(ds)^(m)C_(ds)A_(ds) 1038 ^(m)C_(ds)A_(ds) ^(m)C_(ko)T_(es)G_(es)G_(e)788814 236 252  4687  4703 T_(es) ^(m)C_(eo)^(m)C_(eo)T_(eo)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)A_(ds) ^(m)C_(ds)^(m)C_(ds)  260 A_(ds) ^(m)C_(ds)A_(ko) ^(m)C_(es)T_(es)G_(e) 788815 237253  4688  4704 T_(es)T_(eo) ^(m)C_(eo)^(m)C_(eo)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)A_(ds) ^(m)C_(ds)  335^(m)C_(ds)A_(ds) ^(m)C_(ko)A_(es) ^(m)C_(es)T_(e) 788816 238 254  4689 4705 A_(es)T_(eo)T_(eo) ^(m)C_(eo) ^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds)^(m)C_(ds)A_(ds) ^(m)C_(ds)  412 ^(m)C_(ds)A_(ko) ^(m)C_(es)A_(es)^(m)C_(e) 788817 239 255  4690  4706 A_(es)A_(eo)T_(eo)T_(eo) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)A_(ds)  577 ^(m)C_(ds)^(m)C_(ko)A_(es) ^(m)C_(es)A_(e) 788818 240 256  4691  4707G_(es)A_(eo)A_(eo)T_(eo)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)   33 A_(ds) ^(m)C_(ko)^(m)C_(es)A_(es) ^(m)C_(e) 788819 N/A N/A  4686  4702 ^(m)C_(es)^(m)C_(eo)T_(eo)T_(es)T_(ds)A_(ds) ^(m)C_(ds)A_(ds) ^(m)C_(ds)^(m)C_(ds)A_(ds) 1038 ^(m)C_(ds)A_(ds) ^(m)C_(ko)T_(es)G_(es)G_(e)788820 236 252  4687  4703 T_(es) ^(m)C_(eo)^(m)C_(eo)T_(es)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)A_(ds) ^(m)C_(ds)^(m)C_(ds)  260 A_(ds) ^(m)C_(ds)A_(ko) ^(m)C_(es)T_(es)G_(e) 788821 237253  4688  4704 T_(es)T_(eo) ^(m)C_(eo)^(m)C_(es)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)A_(ds) ^(m)C_(ds)^(m)C_(ds)  335 A_(ds) ^(m)C_(ko)A_(es) ^(m)C_(es)T_(e) 788822 238 254 4689  4705 A_(es)T_(eo)T_(eo) ^(m)C_(es)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)A_(ds) ^(m)C_(ds)  412^(m)C_(ds)A_(ko) ^(m)C_(es)A_(es) ^(m)C_(e) 788823 239 255  4690  4706A_(es)A_(eo)T_(eo)T_(es) ^(m)C_(ds) ^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds)^(m)C_(ds)A_(ds)  577 ^(m)C_(ds) ^(m)C_(ko)A_(es) ^(m)C_(es)A_(e) 788824240 256  4691  4707 G_(es)A_(eo)A_(eo)T_(es)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)   33 A_(ds) ^(m)C_(ko)^(m)C_(es)A_(es) ^(m)C_(e) 788830 236 252  4687  4703 T_(es) ^(m)C_(eo)^(m)C_(es)T_(es)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)A_(ds) ^(m)C_(ds)^(m)C_(ds)A_(ds)  260 ^(m)C_(ds)A_(ko) ^(m)C_(es)T_(es)G_(e) 788831 237253  4688  4704 T_(es)T_(eo) ^(m)C_(es)^(m)C_(es)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)A_(ds) ^(m)C_(ds)^(m)C_(ds)  335 A_(ds) ^(m)C_(ko)A_(es) ^(m)C_(es)T_(e) 788832 238 254 4689  4705 A_(es)T_(eo)T_(es) ^(m)C_(es)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds)A_(ds) ^(m)C_(ds)  412^(m)C_(ds)A_(ko) ^(m)C_(es)A_(es) ^(m)C_(e) 788833 239 255  4690  4706A_(es)A_(eo)T_(es)T_(es) ^(m)C_(ds) ^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds)^(m)C_(ds)A_(ds)  577 ^(m)C_(ds) ^(m)C_(ko)A_(es) ^(m)C_(es)A_(e) 788855N/A N/A 29458 29474T_(es)T_(eo)T_(eo)G_(es)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)T_(ds)2789 ^(m)C_(ko)T_(ks) ^(m)C_(es)T_(e) 788856 N/A N/A 29459 29475T_(es)T_(eo)T_(eo)T_(es)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)T_(ko)2790 ^(m)C_(ks)T_(es) ^(m)C_(e) 788889 N/A N/A 29458 29474T_(es)T_(eo)T_(eo)G_(es)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)T_(ds)2789 ^(m)C_(ko)T_(es) ^(m)C_(es)T_(e) 788890 N/A N/A 29459 29475T_(es)T_(eo)T_(eo)T_(es)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)T_(ko)2790 ^(m)C_(es)T_(es) ^(m)C_(e) 789229 N/A N/A  4686  4705A_(es)T_(eo)T_(eo) ^(m)C_(es) ^(m)C_(es)T_(ds)T_(ds)T_(ds)A_(ds)^(m)C_(ds)A_(ds) ^(m)C_(ds) 1682 ^(m)C_(ds)A_(ds) ^(m)C_(ds)A_(eo)^(m)C_(eo)T_(es)G_(es)G_(e) 789230 237 256  4688  4707G_(es)A_(eo)A_(eo)T_(es)T_(es) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds) 1638 A_(ds) ^(m)C_(ds)^(m)C_(ds)A_(eo) ^(m)C_(eo)A_(es) ^(m)C_(es)T_(e) 789231 238 257  4689 4708 T_(es)G_(eo)A_(eo)A_(es)T_(es)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) 1639 ^(m)C_(ds)A_(ds) ^(m)C_(ds)^(m)C_(eo)A_(eo) ^(m)C_(es)A_(es) ^(m)C_(e) 789232 239 258  4690  4709A_(es)T_(eo)G_(eo)A_(es)A_(es)T_(ds)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) 1640 ^(m)C_(ds)A_(ds) ^(m)C_(eo)^(m)C_(eo)A_(es) ^(m)C_(es)A_(e) 789233 240 259  4691  4710A_(es)A_(eo)T_(eo)G_(es)A_(es)A_(ds)T_(ds)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds) 1641 A_(ds) ^(m)C_(ds)A_(eo) ^(m)C_(eo)^(m)C_(es)A_(es) ^(m)C_(e) 789234 N/A N/A 29457 29476T_(es)T_(eo)T_(eo)T_(es)T_(es)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)1904 T_(ds) ^(m)C_(eo)T_(eo) ^(m)C_(es)T_(es)G_(e) 789235 N/A N/A  4686 4705 A_(es)T_(eo)T_(es) ^(m)C_(es) ^(m)C_(es)T_(ds)T_(ds)T_(ds)A_(ds)^(m)C_(ds)A_(ds) ^(m)C_(ds) 1682 ^(m)C_(ds)A_(ds) ^(m)C_(ds)A_(eo)^(m)C_(eo)T_(es)G_(es)G_(e) 789236 237 256  4688  4707G_(es)A_(eo)A_(es)T_(es)T_(es) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds) 1638 A_(ds) ^(m)C_(ds)^(m)C_(ds)A_(eo) ^(m)C_(eo)A_(es) ^(m)C_(es)T_(e) 789237 238 257  4689 4708 T_(es)G_(eo)A_(es)A_(es)T_(es)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) 1639 ^(m)C_(ds)A_(ds) ^(m)C_(ds)^(m)C_(eo)A_(eo) ^(m)C_(es)A_(es) ^(m)C_(e) 789238 239 258  4690  4709A_(es)T_(eo)G_(es)A_(es)A_(es)T_(ds)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) 1640 ^(m)C_(ds)A_(ds) ^(m)C_(eo)^(m)C_(eo)A_(es) ^(m)C_(es)A_(e) 789239 240 259  4691  4710A_(es)A_(eo)T_(es)G_(es)A_(es)A_(ds)T_(ds)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds) 1641 A_(ds) ^(m)C_(ds)A_(eo) ^(m)C_(eo)^(m)C_(es)A_(es) ^(m)C_(e) 789240 N/A N/A 29457 29476T_(es)T_(eo)T_(es)T_(es)T_(es)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)1904 T_(ds) ^(m)C_(eo)T_(eo) ^(m)C_(es)T_(es)G_(e) 789241 N/A N/A  4686 4705 A_(es)T_(eo)T_(es) ^(m)C_(es) ^(m)C_(es)T_(ds)T_(ds)T_(ds)A_(ds)^(m)C_(ds)A_(ds) ^(m)C_(ds) 1682 ^(m)C_(ds)A_(ds) ^(m)C_(ds)A_(es)^(m)C_(eo)T_(es)G_(es)G_(e) 789242 237 256  4688  4707G_(es)A_(eo)A_(es)T_(es)T_(es) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) ^(m)C_(ds) 1638 A_(ds) ^(m)C_(ds)^(m)C_(ds)A_(es) ^(m)C_(eo)A_(es) ^(m)C_(es)T_(e) 789243 238 257  4689 4708 T_(es)G_(eo)A_(es)A_(es)T_(es)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) 1639 ^(m)C_(ds)A_(ds) ^(m)C_(ds)^(m)C_(es)A_(eo) ^(m)C_(es)A_(es) ^(m)C_(e) 789244 239 258  4690  4709A_(es)T_(eo)G_(es)A_(es)A_(es)T_(ds)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds)A_(ds) 1640 ^(m)C_(ds)A_(ds) ^(m)C_(es)^(m)c_(eo)A_(es) ^(m)c_(es)A_(e) 789245 240 259  4691  4710A_(es)A_(eo)T_(es)G_(es)A_(es)A_(ds)T_(ds)T_(ds) ^(m)C_(ds)^(m)C_(ds)T_(ds)T_(ds)T_(ds) 1641 A_(ds) ^(m)C_(ds)A_(es) ^(m)C_(eo)^(m)C_(es)A_(es) ^(m)C_(e) 789246 N/A N/A 29457 29476T_(es)T_(eo)T_(es)T_(es)T_(es)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)1904 T_(ds)T_(ds) ^(m)C_(es)T_(eo) ^(m)C_(es)T_(es)G_(e) 806693 N/A N/A29458 29477T_(es)T_(eo)T_(es)T_(es)T_(es)T_(ds)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)2791 G_(ds)T_(ds)T_(eo) ^(m)C_(eo)T_(es) ^(m)C_(es)T_(e) 806694 N/A N/A29459 29478T_(es)T_(eo)T_(es)T_(es)T_(es)T_(ds)T_(ds)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)2792 A_(ds)G_(ds)T_(eo)T_(eo) ^(m)C_(es)T_(es) ^(m)C_(e) 806695 N/A N/A29460 29479A_(es)T_(eo)T_(es)T_(es)T_(es)T_(ds)T_(ds)T_(ds)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)2793 T_(ds)A_(ds)G_(eo)T_(eo)T_(es) ^(m)C_(es)T_(e) 806696 N/A N/A 2945429473T_(es)T_(eo)G_(es)T_(es)T_(es)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)T_(ds)^(m)C_(ds) 1901 T_(ds) ^(m)C_(ds)T_(es)G_(eo)T_(es)T_(es)T_(e) 806697N/A N/A 29455 29474T_(es)T_(eo)T_(es)G_(es)T_(es)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)T_(ds)1902 ^(m)C_(ds)T_(ds) ^(m)C_(es)T_(eo)G_(es)T_(es)T_(e) 806698 N/A N/A29456 29475T_(es)T_(eo)T_(es)T_(es)G_(es)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)1903 T_(ds) ^(m)C_(ds)T_(es) ^(m)C_(eo)T_(es)G_(es)T_(e) 806699 N/A N/A29458 29477T_(es)T_(eo)T_(es)T_(es)T_(es)T_(ds)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)2791 G_(ds)T_(ds)T_(eo) ^(m)C_(eo)T_(es) ^(m)C_(es)T_(e) 806700 N/A N/A29459 29478T_(es)T_(eo)T_(es)T_(es)T_(es)T_(ds)T_(ds)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)2792 A_(ds)G_(ds)T_(es)T_(eo) ^(m)C_(es)T_(es) ^(m)C_(e) 806701 N/A N/A29460 29479A_(es)T_(eo)T_(es)T_(es)T_(es)T_(ds)T_(ds)T_(ds)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)2793 T_(ds)A_(ds)G_(es)T_(eo)T_(es) ^(m)C_(es)T_(e) 806708 N/A N/A 2945429473T_(es)T_(eo)G_(eo)T_(es)T_(es)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)T_(ds)^(m)C_(ds) 1901 T_(ds) ^(m)C_(ds)T_(eo)G_(eo)T_(es)T_(es)T_(e) 806709N/A N/A 29455 29474T_(es)T_(eo)T_(eo)G_(es)T_(es)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)T_(ds)1902 ^(m)C_(ds)T_(ds) ^(m)C_(eo)T_(eo)G_(es)T_(es)T_(e) 806710 N/A N/A29456 29475T_(es)T_(eo)T_(eo)T_(es)G_(es)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)1903 T_(ds) ^(m)C_(ds)T_(eo) ^(m)C_(eo)T_(es)G_(es)T_(e) 806711 N/A N/A29458 29477T_(es)T_(eo)T_(eo)T_(es)T_(es)T_(ds)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)2791 G_(ds)T_(ds)T_(es) ^(m)C_(eo)T_(es) ^(m)C_(es)T_(e) 806712 N/A N/A29459 29478T_(es)T_(eo)T_(eo)T_(es)T_(es)T_(ds)T_(ds)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)2792 A_(ds)G_(ds)T_(es)T_(eo) ^(m)C_(es)T_(es) ^(m)C_(e) 806713 N/A N/A29460 29479A_(es)T_(eo)T_(eo)T_(es)T_(es)T_(ds)T_(ds)T_(ds)G_(ds)T_(ds)T_(ds)A_(ds)A_(ds)2793 T_(ds)A_(ds)G_(eo)T_(eo)T_(es) ^(m)C_(es)T_(e) 806714 N/A N/A 2945429473T_(es)T_(eo)G_(es)T_(es)T_(es)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)T_(ds)1901 ^(m)C_(ds)T_(ds) ^(m)C_(ds)T_(eo)G_(eo)T_(es)T_(es)T_(e) 806715 N/AN/A 29455 29474T_(es)T_(eo)T_(es)G_(es)T_(es)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)1902 T_(ds) ^(m)C_(ds)T_(ds) ^(m)C_(eo)T_(eo)G_(es)T_(es)T_(e) 806716N/A N/A 29456 29475T_(es)T_(eo)T_(es)T_(es)G_(es)T_(ds)T_(ds)A_(ds)A_(ds)T_(ds)A_(ds)G_(ds)T_(ds)1903 T_(ds) ^(m)C_(ds)T_(eo) ^(m)C_(es)T_(es)G_(es)T_(e) 827592 N/A N/A19633 19652 A_(es)^(m)C_(eo)A_(eo)G_(es)A_(es)T_(ds)A_(ds)T_(ds)T_(ds)T_(ds)T_(ds)T_(ds)G_(ds)1703 T_(ds)T_(ds) ^(m)C_(eo)T_(eo)G_(es) ^(m)C_(es) ^(m)C_(e) 827599 N/AN/A 19633 19652 A_(es)^(m)C_(eo)A_(es)G_(es)A_(es)T_(ds)A_(ds)T_(ds)T_(ds)T_(ds)T_(ds)T_(ds)G_(ds)1703 T_(ds)T_(ds) ^(m)C_(eo)T_(eo)G_(es) ^(m)C_(es) ^(m)C_(e) 827606 N/AN/A 19633 19652 A_(es)^(m)C_(eo)A_(es)G_(es)A_(es)T_(ds)A_(ds)T_(ds)T_(ds)T_(ds)T_(ds)T_(ds)G_(ds)1703 T_(ds)T_(ds) ^(m)C_(es)T_(eo)G_(es) ^(m)C_(es) ^(m)C_(e) 827607 N/AN/A 21224 21243 G_(es)T_(eo)T_(es)G_(es)T_(es)T_(ds)^(m)C_(ds)A_(ds)G_(ds)A_(ds)A_(ds)T_(ds)T_(ds) 1754A_(ds)T_(ds)G_(eo)T_(eo) ^(m)C_(es)A_(es)T_(e) 827611 N/A N/A 2328623305 ^(m)C_(es)A_(eo)T_(es)A_(es)T_(es)T_(ds)G_(ds)T_(ds)T_(ds)^(m)C_(ds)T_(ds) ^(m)C_(ds) 1804 A_(ds)G_(ds)A_(ds)G_(eo)A_(eo)^(m)C_(es) ^(m)C_(es)A_(e) 827617 N/A N/A 28456 28475^(m)C_(es)A_(eo)G_(es)A_(es)^(m)C_(es)T_(ds)G_(ds)T_(ds)A_(ds)A_(ds)T_(ds) ^(m)C_(ds) 1887T_(ds)A_(ds)G_(ds)G_(eo)A_(eo) ^(m)C_(es) ^(m)C_(es) ^(m)C_(e) 827630N/A N/A 50920 50939 G_(es)T_(eo)T_(es)T_(es)T_(es)^(m)C_(ds)A_(ds)T_(ds) ^(m)C_(ds)A_(ds)A_(ds)T_(ds)A_(ds) 2193 T_(ds)^(m)C_(ds)T_(eo)G_(eo) ^(m)C_(es)A_(es)A_(e) 827649 N/A N/A 21224 21243G_(es)T_(eo)T_(es)G_(es)T_(es)T_(ds)^(m)C_(ds)A_(ds)G_(ds)A_(ds)A_(ds)T_(ds)T_(ds) 1754A_(ds)T_(ds)G_(es)T_(eo) ^(m)C_(es)A_(es)T_(e) 827653 N/A N/A 2328623305 ^(m)C_(es)A_(eo)T_(es)A_(es)T_(es)T_(ds)G_(ds)T_(ds)T_(ds)^(m)C_(ds)T_(ds) ^(m)C_(ds) 1804 A_(ds)G_(ds)A_(ds)G_(es)A_(eo)^(m)C_(es) ^(m)C_(es)A_(e) 827691 N/A N/A 21224 21243G_(es)T_(eo)T_(eo)G_(es)T_(es)T_(ds)^(m)C_(ds)A_(ds)G_(ds)A_(ds)A_(ds)T_(ds)T_(ds) 1754A_(ds)T_(ds)G_(eo)T_(eo) ^(m)C_(es)A_(es)T_(e) 827695 N/A N/A 2328623305 ^(m)C_(es)A_(eo)T_(eo)A_(es)T_(es)T_(ds)G_(ds)T_(ds)T_(ds)^(m)C_(ds)T_(ds) ^(m)C_(ds) 1804 A_(ds)G_(ds)A_(ds)G_(eo)A_(eo)^(m)C_(es) ^(m)C_(es)A_(e) 827701 N/A N/A 28456 28475^(m)C_(es)A_(eo)G_(eo)A_(es)^(m)C_(es)T_(ds)G_(ds)T_(ds)A_(ds)A_(ds)T_(ds) ^(m)C_(ds) 1887T_(ds)A_(ds)G_(ds)G_(eo)A_(eo) ^(m)C_(es) ^(m)C_(es) ^(m)C_(e) 827714N/A N/A 50920 50939 G_(es)T_(eo)T_(eo)T_(es)T_(es)^(m)C_(ds)A_(ds)T_(ds) ^(m)C_(ds)A_(ds)A_(ds)T_(ds)A_(ds) 2193 T_(ds)^(m)C_(ds)T_(eo)G_(eo) ^(m)C_(es)A_(es)A_(e)

Example 7: Effect of Modified Oligonucleotides on Human SNCA In Vitro,Multiple Doses

Modified oligonucleotides selected from the examples above were testedat various doses in SH-SY5Y cells. Comparator oligonucleotide 387978 wasalso tested. Cells were plated at a density of 20,000 cells per well andtransfected using electroporation with 0.55 μM, 1.67 μM, 5.00 μM and15.00 μM concentrations of modified oligonucleotide, as specified in thetables below. After a treatment period of approximately 24 hours, totalRNA was isolated from the cells and SNCA mRNA levels were measured byquantitative real-time PCR Human SNCA primer probe set RTS2621(described hereinabove in Example 1) was used to measure mRNA levels.SNCA mRNA levels were adjusted according to total RNA content, asmeasured by RIBOGREEN®. Results are presented in the tables below aspercent reduction of the amount of SNCA mRNA, relative to untreatedcontrol. A value of 0% reduction indicates that the compound had noeffect or increased mRNA concentrations in the cell. As illustrated inthe tables below, SNCA mRNA levels were reduced in a dose-dependentmanner in modified oligonucleotide-treated cells. IC50 was calculatedusing the “log(inhibitor) vs. response—variable slope (4 parameters)”formula using Prism6 software.

TABLE 46 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound 15.00 IC₅₀Number 0.55 μM 1.67 μM 5.00 μM μM (μM) 387978 20 44 50 75 3.49 709533 1951 59 87 2.27 709551 20 41 61 75 2.97 709556 16 30 62 82 3.34 709654 1647 64 83 2.42 709581 12 31 61 83 3.41 709582 10 34 57 76 3.86 709640 322 54 80 4.62 709875 22 45 58 82 2.60 709882 33 41 50 86 2.57 709893 4370 87 92 0.72 709900 28 50 58 75 2.31 709919 26 55 52 88 2.07 709924 1236 56 76 3.77 709936 16 39 60 80 3.06 709944 42 72 81 90 0.71 709949 2335 55 84 3.16 709967 35 65 62 81 1.17 709979 30 56 67 76 1.62

TABLE 47 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.55μM 1.67 μM 5.00 μM 15.00 μM (μM) 387978 14 19 57 76 4.58 709543 15 37 6175 3.35 709544 33 47 71 87 1.61 709550 33 51 53 78 2.12 709573 26 48 6482 2.08 709579 0 29 67 90 3.19 709580 19 8 58 86 4.49 709856 0 21 48 627.15 709592 1 24 51 73 5.29 709597 7 40 62 86 2.96 709651 17 26 45 715.65 709873 7 24 74 73 3.40 709891 19 40 62 80 2.82 709892 19 33 67 773.01 709897 27 40 55 86 2.67 709898 32 64 87 90 1.04 709909 0 23 59 724.70 709953 0 34 55 75 4.29 709966 23 56 78 87 1.51

TABLE 48 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.55μM 1.67 μM 5.00 μM 15.00 μM (μM) 387978 6 42 65 72 3.23 709536 50 68 8386 <0.55 709548 18 37 59 76 3.33 709549 41 66 82 89 0.81 709555 20 45 5358 4.93 709560 5 37 60 66 4.28 709572 43 54 64 79 1.12 709578 16 37 5679 3.44 709591 0 23 50 86 4.64 709596 0 19 51 70 5.83 709632 14 39 65 772.97 709668 0 0 8 27 29.96 709788 0 12 40 72 7.11 709878 26 71 78 931.09 709879 17 50 84 93 1.66 709890 41 53 72 87 1.13 709915 37 64 65 861.07 709940 14 36 67 84 2.80 709977 37 34 49 68 4.04

TABLE 49 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.55μM 1.67 μM 5.00 μM 15.00 μM (μM) 387978 11 30 62 72 3.92 709534 23 48 6781 2.11 709535 44 80 74 79 <0.55 709547 37 55 61 83 1.42 709558 27 56 5656 3.29 709563 12 49 64 77 2.61 709565 25 60 68 78 1.62 709571 9 47 5969 3.45 709576 12 37 69 68 3.28 709588 9 53 84 84 1.73 709625 0 24 47 765.44 709727 0 34 49 71 5.14 709751 0 8 27 57 11.73 709877 27 52 75 881.56 709881 18 29 58 72 4.10 709883 21 56 76 83 1.63 709894 25 37 76 902.08 709895 24 49 77 91 1.67 709912 4 36 54 73 4.32

Example 8: Effect of Modified Oligonucleotides on Human SNCA In Vitro,Multiple Doses

Modified oligonucleotides selected from the examples above were testedat various doses in SH-SY5Y cells. Cells were plated at a density of20,000 cells per well and transfected using electroporation with 0.48μM, 1.44 μM, 4.33 μM, and 13.00 μM concentrations of modifiedoligonucleotide, as specified in the tables below. After a treatmentperiod of approximately 24 hours, total RNA was isolated from the cellsand SNCA mRNA levels were measured by quantitative real-time PCR HumanSNCA primer probe set RTS2621 (described hereinabove in Example 1) wasused to measure mRNA levels. SNCA mRNA levels were adjusted according tototal RNA content, as measured by RIBOGREEN®. Results are presented inthe tables below as percent reduction of the amount of SNCA mRNA,relative to untreated control. A value of 0% reduction indicates thatthe compound had no effect or increased mRNA concentrations in the cell.As illustrated in the tables below, SNCA mRNA levels were reduced in adose-dependent manner in modified oligonucleotide-treated cells.

TABLE 50 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.48μM 1.44 μM 4.33 μM 13.00 μM (μM) 740363 7 37 64 89 2.53 740364 28 35 6383 2.3 740406 45 66 85 83 0.59 740407 46 79 84 93 0.51 740408 68 81 9395 <0.48 740409 49 70 92 93 0.52 740410 43 70 80 90 0.62 740411 19 63 8691 1.14 740412 31 68 75 93 0.92 740416 41 63 84 94 0.74 740422 51 69 8686 0.43 740425 62 75 78 93 n/a 740427 45 75 87 63 <0.48 740429 33 56 7286 1.16 740430 42 70 90 95 0.65 740432 0 0 0 17 >13 740438 32 54 67 891.29 740439 39 61 79 96 0.84 740440 36 55 79 94 1.01

TABLE 51 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.48μM 1.44 μM 4.33 μM 13.00 μM (μM) 740410 43 59 76 68 0.68 740414 29 44 6971 1.91 740431 51 63 68 72 <0.48 740456 39 51 47 70 2.02 740482 30 44 6370 2.14 740498 47 45 68 74 1.02 740500 46 53 79 78 0.74 740508 10 38 5570 3.63 740509 44 67 80 85 0.6 740510 24 54 69 83 1.56 740513 19 35 6644 6.31 740517 5 38 73 61 3.07 740527 0 47 65 80 2.43 740528 11 45 77 831.88 740533 12 29 76 81 2.43 740534 21 56 40 5 n/a 740535 7 37 24 629.22 740545 32 58 69 81 1.2 740612 0 16 69 65 4.02

TABLE 52 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.48μM 1.44 μM 4.33 μM 13.00 μM (μM) 740140 45 58 69 82 0.75 740585 19 46 6662 2.70 740601 15 47 61 63 3.05 740604 16 34 45 62 5.64 740608 2 15 5963 5.13 740610 8 43 58 56 4.49 740615 32 50 59 76 1.81 740625 7 33 49 783.88 740649 7 25 54 41 11.78 740650 25 34 54 66 3.80 740654 27 32 50 753.45 740668 36 41 53 44 >13 740670 0 40 45 32 >13 740692 21 28 52 654.64 740730 42 47 62 78 1.30 740783 36 48 48 74 2.23 740794 33 35 5337 >13 740801 10 38 58 67 3.62 740802 19 12 55 63 5.73

TABLE 53 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.48μM 1.44 μM 4.33 μM 13.00 μM (μM) 740140 41 52 71 68 1.04 740796 3 28 5154 6.89 740808 9 32 47 73 4.40 740851 2 21 46 74 5.05 740893 4 24 63 723.71 740894 0 20 46 33 >13 740904 6 19 51 72 4.81 740919 23 43 68 901.84 740922 0 23 49 63 5.89 740923 0 29 52 79 3.93 740927 8 29 58 823.30 740976 16 44 65 72 2.52 740997 0 26 61 74 3.69 741000 25 52 43 683.43 741001 34 52 65 78 1.41 741002 14 37 62 69 3.19 741005 6 19 45 656.14 741006 54 76 71 78 <0.48 741008 42 54 72 80 0.92

TABLE 54 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.48μM 1.44 μM 4.33 μM 13.00 μM (μM) 740140 35 60 71 81 1.03 740939 13 28 6080 3.21 740941 8 30 38 83 4.55 740972 0 29 64 73 3.43 741009 0 32 52 833.62 741321 59 69 79 89 <0.48 741378 21 49 84 86 1.43 741410 57 79 85 92<0.48 741455 53 69 88 87 <0.48 741472 11 40 61 60 3.83 741473 9 29 53 654.76 741477 32 32 50 49 10.65 741483 11 23 53 62 5.42 741487 19 39 72 702.36 741502 14 26 51 61 5.58 741514 0 27 41 73 5.34 741516 8 13 63 704.29 741525 9 32 58 61 4.70 741540 0 25 41 26 <0.48

Example 9: Effect of Modified Oligonucleotides on Human SNCA In Vitro,Multiple Doses

Modified oligonucleotides selected from the examples above were testedat various doses in SH-SY5Y cells. Comparator oligonucleotide 397978 wasalso tested. Cells were plated at a density of 20,000 cells per well andtransfected using electroporation with 0.11 μM, 0.33 μM, 1.00 μM, and3.00 μM concentrations of modified oligonucleotide, as specified in thetables below. After a treatment period of approximately 24 hours, totalRNA was isolated from the cells and SNCA mRNA levels were measured byquantitative real-time PCR. Human SNCA primer probe set RTS2621(described hereinabove in Example 1) was used to measure mRNA levels.SNCA mRNA levels were adjusted according to total RNA content, asmeasured by RIBOGREEN®. Results are presented in the tables below aspercent reduction of the amount of SNCA mRNA, relative to untreatedcontrol. A value of 0% reduction indicates that the compound had noeffect or increased mRNA concentrations in the cell. As illustrated inthe tables below, SNCA mRNA levels were reduced in a dose-dependentmanner in modified oligonucleotide-treated cells.

TABLE 55 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.11μM 0.33 μM 1.00 μM 3.00 μM (μM) 740140 16 43 62 90 0.51 741021 19 56 8193 0.30 741022 28 62 83 87 0.24 741028 28 54 78 94 0.28 741029 36 57 8783 0.22 741032 23 56 86 93 0.28 741037 26 64 88 89 0.23 741077 34 53 8489 0.24 741122 30 69 88 87 0.20 741125 25 57 83 91 0.27 741169 34 66 9496 0.19 741170 34 71 92 93 0.18 741189 35 56 81 95 0.23 741206 32 57 8194 0.24 741207 26 63 91 96 0.23 741228 23 49 78 90 0.34 741229 10 58 8195 0.31 741278 28 49 76 92 0.31 741379 22 70 86 93 0.22

TABLE 56 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.11μM 0.33 μM 1.00 μM 3.00 μM (μM) 740140 13 38 70 87 0.50 741016 30 42 7491 0.36 741018 10 32 65 88 0.61 741019 14 20 56 86 0.82 741030 17 27 6489 0.63 741034 12 28 47 80 0.93 741038 14 42 72 91 0.46 741039 63 42 7093 0.11 741043 13 33 66 83 0.60 741047 29 45 72 94 0.35 741049 18 34 5884 0.65 741073 4 46 75 93 0.43 741078 23 52 72 92 0.33 741080 15 37 6678 0.59 741082 18 48 75 95 0.37 741083 17 36 72 82 0.51 741101 18 44 7992 0.38 741111 0 29 68 88 0.64 741129 5 38 54 77 0.79

TABLE 57 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.11 μM0.33 μM 1.00 μM 3.00 μM IC₅₀ (μM) 740140 39 34 69 81 0.39 741094 3 13 5678 0.99 741110 5 26 47 66 1.26 741113 27 26 69 76 0.60 741167 16 37 7079 0.54 741168 23 39 63 84 0.52 741178 29 34 58 77 0.62 741179 22 54 7189 0.34 741188 0 25 70 77 0.70 741190 18 51 70 78 0.41 741191 23 37 6382 0.55 741195 0 20 58 81 0.88 741197 20 34 69 76 0.58 741201 6 25 46 811.01 741205 14 34 66 87 0.57 741208 13 51 72 82 0.42 741227 8 30 61 780.74 741230 10 27 45 73 1.10 741231 0 20 33 82 1.31

TABLE 58 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.11 μM0.33 μM 1.00 μM 3.00 μM IC₅₀ (μM) 387978 11 0 7 39 >3.0 740140 26 35 5280 0.67 740432 19 36 53 74 0.77 741187 18 30 60 78 0.72 741214 7 18 5279 1.00 741220 16 35 49 73 0.91 741234 20 41 71 85 0.45 741241 15 25 5378 0.88 741246 3 26 54 68 1.06 741280 21 44 62 78 0.52 741301 22 46 6684 0.44 741315 25 46 68 83 0.40 741320 17 36 65 85 0.55 741329 24 40 6182 0.53 741330 13 36 69 85 0.55 741335 12 35 60 76 0.72 741368 18 46 7287 0.42 741373 10 31 58 73 0.83 741393 17 37 67 82 0.54

Example 10: Effect of Modified Oligonucleotides on Human SNCA In Vitro,Multiple Doses

Modified oligonucleotides selected from the examples above were testedat various doses in SH-SY5Y cells. Compound No. 387985, previouslydisclosed in WO 2012/068405 was also tested and is comparatoroligonucleotide. Cells were plated at a density of 20,000 cells per welland transfected using electroporation with 0.44 μM, 1.33 μM, 4.00 μM,and 12.00 μM concentrations of modified oligonucleotide, as specified inthe tables below. After a treatment period of approximately 24 hours,total RNA was isolated from the cells and SNCA mRNA levels were measuredby quantitative real-time PCR Human SNCA primer probe set RTS2621(described hereinabove in Example 1) was used to measure mRNA levels.SNCA mRNA levels were adjusted according to total RNA content, asmeasured by RIBOGREEN®. Results are presented in the tables below aspercent reduction of the amount of SNCA mRNA, relative to untreatedcontrol. A value of 0% reduction indicates that the compound had noeffect or increased mRNA concentrations in the cell. As illustrated inthe tables below, SNCA mRNA levels were reduced in a dose-dependentmanner in modified oligonucleotide-treated cells.

TABLE 59 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.44 μM1.33 μM 4.00 μM 12.00 μM IC₅₀ (μM) 740140 42 76 86 93 0.56 741140 77 7496 86 <0.44 762851 24 45 76 93 1.45 762855 16 45 85 90 1.46 762858 25 5584 96 1.11 762880 9 33 55 85 2.93 762882 75 92 94 90 0.07 762886 39 6782 84 0.66 762891 25 61 88 93 0.97 762893 31 63 86 95 0.87 762899 42 7488 82 0.53 762900 69 93 96 97 <0.44 762901 63 86 90 93 <0.44 762924 3059 82 84 0.97 762926 40 63 80 92 0.72 762930 37 73 85 80 0.62 762932 4272 94 93 0.58 762953 39 61 85 91 0.76 762969 60 72 86 92 <0.44

TABLE 60 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.44 μM1.33 μM 4.00 μM 12.00 μM IC₅₀ (μM) 740140 58 79 70 75 <0.44 741140 84 9389 79 >12 762914 80 87 91 95 <0.44 762952 42 74 85 94 0.57 762960 34 5479 82 1.01 762962 52 81 84 94 <0.44 762965 48 72 91 93 0.48 762987 37 6986 75 0.65 763002 37 54 79 88 0.94 763019 30 51 77 91 1.19 763032 47 7084 78 <0.44 763033 68 84 89 86 <0.44 763035 40 69 85 91 0.64 763040 3667 85 94 0.73 763049 34 68 82 88 0.77 763050 54 87 87 84 <0.44 763052 5475 85 91 <0.44 763059 4 67 83 89 1.08 763087 51 93 91 94 <0.44

TABLE 61 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.44 μM1.33 μM 4.00 μM 12.00 μM IC₅₀ (μM) 740140 48 60 61 75 n/a 741140 80 8477 93 n/a 763065 22 55 73 89 1.32 763072 45 66 76 91 0.57 763075 39 5886 89 0.78 763079 49 74 86 83 <0.44 763081 39 69 70 83 0.68 763084 50 7375 89 <0.44 763085 57 81 88 83 <0.44 763088 73 72 79 89 <0.44 763102 2055 77 68 1.51 763142 58 70 79 87 <0.44 763150 44 60 74 84 0.68 763151 3971 68 82 0.64 763166 52 77 75 84 <0.44 763177 60 64 89 72 <0.44 76318222 57 67 76 1.49 763196 52 68 86 83 <0.44 763207 49 73 79 82 <0.44

TABLE 62 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.44 μM1.33 μM 4.00 μM 12.00 μM IC₅₀ (μM) 740140 48 66 84 82 0.45 741140 77 8889 97 <0.44 763145 79 81 74 92 <0.44 763185 31 64 79 86 0.90 763188 3670 85 90 0.69 763216 41 72 68 94 0.61 763225 54 74 84 93 <0.44 763228 1853 79 93 1.32 763233 45 68 83 94 0.56 763249 25 48 65 89 1.56 763272 3860 89 95 0.77 763281 58 74 92 96 <0.44 763291 27 35 67 86 1.92 763299 5271 83 87 <0.44 763309 43 43 72 76 1.11 763312 45 68 82 94 0.56 763333 1255 85 81 1.32 763341 34 61 67 89 0.96 763364 51 74 72 90 <0.44

TABLE 63 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.44 μM1.33 μM 4.00 μM 12.00 μM IC₅₀ (μM) 740140 27 47 55 55 3.65 741140 48 5656 49 <0.44 763295 22 51 64 71 1.88 763305 28 52 54 71 2.05 763348 27 4051 68 3.12 763355 43 49 51 62 1.77 763359 35 57 61 68 1.32 763360 49 5571 74 0.55 763370 18 54 61 75 1.93 763373 44 45 69 70 1.09 763384 23 5240 69 3.43 763391 28 48 55 66 2.49 763393 49 36 50 71 1.96 763394 25 4171 86 1.68 763418 0 31 57 41 9.00 763434 38 66 50 65 1.04 763440 25 4649 51 <0.44 763481 34 51 57 69 1.74 763491 32 47 54 64 2.53

TABLE 64 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.44 μM1.33 μM 4.00 μM 12.00 μM IC₅₀ (μM) 740140 47 59 54 50 <0.44 741140 62 5874 68 <0.44 763466 19 47 55 66 2.87 763485 34 49 71 70 1.33 763492 7 4440 62 5.24 763501 25 36 59 62 3.35 763612 33 44 45 71 2.76 763627 39 6260 68 0.91 763628 22 48 42 57 <0.44 763629 31 48 53 76 2.00 763634 26 4144 57 5.83 763640 12 44 31 74 4.70 763641 6 33 51 76 3.58 763643 16 3249 57 5.74 763645 18 36 55 71 3.22 763650 45 61 46 65 0.83 763651 43 6064 59 0.62 763671 17 47 43 59 5.08 763741 23 37 61 71 2.64

TABLE 65 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.44 μM1.33 μM 4.00 μM 12.00 μM IC₅₀ (μM) 740140 42 61 84 75 0.65 741140 72 7992 94 <0.44  762837* 20 37 62 81 2.4 763684 13 44 75 92 1.69 763685 2764 83 90 0.95 763693 3 15 58 76 3.88 763701 1 38 74 77 2.26 763702 16 3768 84 2.15 763706 12 48 57 86 2.17 763712 18 50 56 89 1.90 763721 1 3761 60 3.90 763723 19 42 65 77 2.16 763734 0 22 56 77 3.78 763740 32 4777 88 1.23 763749 24 61 81 71 1.16 763755 8 45 57 72 2.82 763778 25 5978 76 1.18 763793 41 71 81 93 0.61 763823 23 60 82 71 1.17 *Valuesrepresent the average of three experiments

TABLE 66 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides in SH-SY5Y cells SNCA inhibition (% reduction) CompoundNumber 0.44 μM 1.33 μM 4.00 μM 12.00 μM IC₅₀ (μM) 740140 50 59 68 930.58 741140 75 64 83 88 n/a 762837 12 37 64 82 2.43 762837 27 33 51 862.64 763795 17 42 62 79 2.26 763798 25 43 63 78 2.00 763817 10 38 57 882.55 763818 24 53 52 92 1.72 763829 41 68 66 94 0.68 763876 33 65 83 840.81 763877 25 49 47 83 2.23 763882 27 57 67 86 1.27 763883 44 63 71 920.67 763884 25 42 64 78 2.00 763936 19 31 56 80 2.92 763939 18 27 54 654.27 763949 12 28 58 67 3.83 763972 0 36 46 67 4.60 763985 6 28 60 653.86

TABLE 67 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.44 μM1.33 μM 4.00 μM 12.00 μM IC₅₀ (μM) 740140 41 60 74 82 0.73 741140 79 8991 87 <0.44 763295 8 38 77 91 1.88 763305 19 52 75 89 1.41 763348 28 6278 93 0.97 763355 25 61 86 85 0.99 763359 49 76 90 94 0.45 763360 60 8694 94 <0.44 763370 28 68 89 90 0.83 763373 51 59 79 90 0.51 763384 37 6785 92 0.70 763391 30 64 79 82 0.94 763393 35 76 84 93 0.67 763394 36 7683 96 0.65 763418 0 26 54 81 3.52 763434 40 73 91 92 0.60 763440 15 5174 83 1.57 763481 35 73 84 88 0.68 763491 31 53 80 83 1.11

TABLE 68 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound Number 0.44 μM1.33 μM 4.00 μM 12.00 μM IC₅₀ (μM) 740140 36 53 69 87 1.08 741140 60 7087 88 <0.44 763466 27 52 74 80 1.34 763485 36 66 80 87 0.75 763492 9 4171 87 1.97 763501 24 48 71 79 1.58 763612 37 63 82 90 0.76 763627 41 6384 82 0.66 763628 44 72 85 86 0.53 763629 29 59 80 85 1.02 763634 22 5581 92 1.18 763640 34 48 80 88 1.13 763641 21 53 79 82 1.33 763643 35 5382 82 0.97 763645 0 55 65 84 1.93 763650 46 71 81 92 0.51 763651 40 5782 86 0.79 763671 0 31 54 76 3.55 763741 32 40 71 73 1.71

TABLE 69 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) IC₅₀ Number 0.44μM 1.33 μM 4.00 μM 12.00 μM (μM) 387985 0 22 48 67 5.00 789243 9 38 6679 2.40 827599 52 71 86 91 0.40

Example 11: Effect of Modified Oligonucleotides on Human SNCA In Vitro,Multiple Doses

Modified oligonucleotides selected from the examples above were testedat various doses in SH-SY5Y cells. Compound No. 387985, previouslydisclosed in WO 2012/068405 was also tested and is comparatoroligonucleotide. Cells were plated at a density of 20,000 cells per welland transfected using electroporation with 0.032 μM, 0.160 μM, 0.800 μM,4.000 μM, and 20.000 μM concentrations of modified oligonucleotide, asspecified in the tables below. After a treatment period of approximately24 hours, total RNA was isolated from the cells and SNCA mRNA levelswere measured by quantitative real-time PCR Human SNCA primer probe setRTS2621 (described hereinabove in Example 1) was used to measure mRNAlevels. SNCA mRNA levels were adjusted according to total RNA content,as measured by RIBOGREEN®. Results are presented in the tables below aspercent reduction of the amount of SNCA mRNA, relative to untreatedcontrol. A value of 0% reduction indicates that the compound had noeffect or increased mRNA concentrations in the cell. As illustrated inthe tables below, SNCA mRNA levels were reduced in a dose-dependentmanner in modified oligonucleotide-treated cells.

TABLE 70 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) Number 0.032 μM0.160 μM 0.800 μM 4.000 μM 20.000 μM IC₅₀ (μM) 762837 0 7 33 76.1 911.72 762901 8 33 73 91 93 0.32 762952 0 15 67 81 95 0.57 763002 0 8 4775 94 1.10 763032 11 35 71 77 96 0.36 763085 0 31 63 83 94 0.47 763364 016 51 81 89 0.86 763391 0 14 48 71 91 1.10 788833 0 8 39 77 90 1.31789239 0 0 14 61 87 3.02 789242 0 1 27 60 85 2.67 789243 0 1 27 64 902.25

TABLE 71 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides SNCA inhibition (% reduction) Compound 0.032 0.1600.800 4.000 20.000 IC₅₀ Number μM μM μM μM μM (μM) 387985 0 0 13 54 774.20 789243 0 7 31 69 85 1.90 827599 10 34 65 84 91 0.40

Example 12: Effect of Modified Oligonucleotides on Human SNCA In Vitro,Multiple Doses

Modified oligonucleotides selected from the examples above were testedat various doses in A431 cells. Cells were plated at a density of 5,000cells per well and transfected by free uptake with 0.032 μM, 0.160 μM,0.800 μM, 4.000 μM, and 20.000 μM concentrations of modifiedoligonucleotide, as specified in the tables below. After a treatmentperiod of approximately 24 hours, total RNA was isolated from the cellsand SNCA mRNA levels were measured by quantitative real-time PCR HumanSNCA primer probe set RTS2621 (described hereinabove in Example 1) wasused to measure mRNA levels. SNCA mRNA levels were adjusted according tototal RNA content, as measured by RIBOGREEN®. Results are presented inthe tables below as percent reduction of the amount of SNCA mRNA,relative to untreated control. A value of 0% reduction indicates thatthe compound had no effect or increased mRNA concentrations in the cell.As illustrated in the tables below, SNCA mRNA levels were reduced in adose-dependent manner in modified oligonucleotide-treated cells.

TABLE 72 Dose-dependent percent reduction of human SNCA mRNA by modifiedoligonucleotides Compound SNCA inhibition (% reduction) Number 0.032 μM0.160 μM 0.800 μM 4.000 μM 20.000 μM IC₅₀ (μM) 762837 0 6 7 33 51 16.95762901 1 28 53 60 81 1.20 762952 16 38 59 81 93 0.41 763002 0 6 41 71 871.49 763032 12 60 89 96 97 0.13 763085 2 19 75 90 96 0.40 763364 38 5777 93 97 0.09 763391 5 28 71 92 95 0.37 788833 8 23 53 71 93 0.82 7892396 0 32 32 45 0.00 789242 0 0 4 32 66 9.78 789243 0 3 13 39 66 8.02

Example 13: Effect of Modified Oligonucleotides on Rhesus Monkey SNCA InVitro, Multiple Doses

Several of the modified oligonucleotides described hereinabove arecomplementary to rhesus monkey Human-monkey cross reactive modifiedoligonucleotides selected from the examples above were tested at variousdoses in LLC-MK2 monkey cells. Cells were plated at a density of 20,000cells per well and transfected using electroporation with 6.9 nM, 20.5nM, 61.8 nM, 185.2 nM, 500.0 nM, 1700.0 nM, 5000.0 nM, and 15,000.0 nMconcentrations of modified oligonucleotide, as specified in the tablebelow. After a treatment period of approximately 24 hours, total RNA wasisolated from the cells and SNCA mRNA levels were measured byquantitative real-time PCR Human SNCA primer probe set RTS2621(described hereinabove in Example 1) was used to measure mRNA levels.SNCA mRNA levels were adjusted according to total RNA content, asmeasured by RIBOGREEN®. Results are presented in the tables below aspercent reduction of the amount of SNCA mRNA, relative to untreatedcontrol. The half maximal inhibitory concentration (IC₅₀) of eacholigonucleotide is also presented in the table below. A value of 0%reduction indicates that the compound had no effect or increased mRNAconcentrations in the cell. As illustrated in the tables below, SNCAmRNA levels were reduced in a dose-dependent manner in modifiedoligonucleotide-treated cells.

TABLE 73 Dose-dependent percent reduction of rhesus monkey SNCA mRNA bymodified oligonucleotides SNCA Inhibition (% Reduction) Compound 6.920.5 61.8 185.2 500.0 1700.0 5000.0 15,000.0 IC₅₀ No. nM nM nM nM nM nMnM nM (μM) 709534 0 0 0 0 8 46 66 79 2.8 709535 0 0 0 7 28 59 84 90 1.4709536 0 0 0 16 44 66 78 90 1.1 709883 0 0 0 0 36 61 76 88 1.5 709967 00 0 0 8 35 73 90 2.5  741082* 0 0 0 0 0 0 36 78 7.0 *one mismatch torhesus monkey

Example 14: Effect of Modified Oligonucleotides on Monkey SNCA In Vitro,Multiple Doses

Several of the modified oligonucleotides described hereinabove arecomplementary to rhesus monkeys. Human-monkey cross reactive modifiedoligonucleotides selected from the examples above were tested at variousdoses in LLC-MK2 monkey cells. Modified oligonucleotides with 1-3mismatches to rhesus monkey sequence are marked in the table below.Cells were plated at a density of 20,000 cells per well and transfectedusing electroporation with 0.032 μM, 0.160 μM, 0.800 μM, 4.000 μM, and20.000 μM concentrations of modified oligonucleotide, as specified inthe table below. After a treatment period of approximately 24 hours,total RNA was isolated from the cells and SNCA mRNA levels were measuredby quantitative real-time PCR Human SNCA primer probe set RTS2621(described hereinabove in Example 1) was used to measure mRNA levels.SNCA mRNA levels were adjusted according to total RNA content, asmeasured by RIBOGREEN®. Results are presented in the tables below aspercent reduction of the amount of SNCA mRNA, relative to untreatedcontrol. As illustrated in the tables below, SNCA mRNA levels werereduced in a dose-dependent manner in modified oligonucleotide-treatedcells.

TABLE 74 Dose-dependent percent reduction of rhesus monkey SNCA mRNA bymodified oligonucleotides Compound SNCA inhibition (% reduction) Number0.032 μM 0.160 μM 0.800 μM 4.000 μM 20.000 μM IC₅₀ (μM) 762837 0 0 5 5784 3.67 762901 0 18 47 88 98 0.80  762952** 0 0 0 14 53 17.90 763002 120 36 85 93 1.23 763032 6 29 63 85 94 0.47  763085* 0 0 28 60 81 2.75 763364* 0 0 28 58 85 2.77 763391 0 3 50 65 87 1.46 788833 0 0 15 59 843.30 789239 0 0 3 45 78 5.61 789242 0 0 5 41 78 6.05 789243 0 0 19 56 853.28 *one mismatch to monkey; **two mismatches to monkey

Example 15: Effect of Modified Oligonucleotides on Human SNCA in HumanNeurons by Free Uptake, Single Dose

Selected modified oligonucleotides complementary to human SNCA weretested for their effects on SCNA mRNA levels in human neurons in vitroby free uptake. Human IPS-cell derived neurons were plated at a densityof 35,000 cells per well. After approximately 24 hours, 20 μM modifiedoligonucleotide was added and incubated with the cultured cells for 7days. After 7 days, total RNA was isolated from the cells and SNCA mRNAlevels were measured by quantitative real-time PCR Human SNCA primerprobe set RTS2621 (described hereinabove in Example 1) was used tomeasure mRNA levels. SNCA mRNA levels were adjusted according to totalRNA content, as measured by RIBOGREEN®. Results are presented in thetables below as percent reduction of the amount of SNCA mRNA relative tountreated control cells. As shown below, modified oligonucleotidescomplementary to human SNCA reduced the amount of human SNCA mRNA.

TABLE 75 Percent reduction of human SNCA mRNA in human neurons by freeuptake Compound No % Reduction 709897 53 740425 58 741082 98 762836 80762837 79 762838 75 762839 64 762840 54 762895 60 762896 63 762898 82762899 86 762900 85 762901 41 762914 74 762948 33 762949 96 762951 45762952 77 763001 84 763002 82 763003 86 763004 82 763032 88 763033 82763035 68 763040 71 763050 78 763084 89 763085 95 763087 78 763102 66763196 56 763207 90 763216 65 763233 68 763364 83 763391 95 763393 81763813 57 763817 51 763818 61 788815 63 788816 67 788820 63 788821 69788822 76 788823 81 788824 73 788830 64 788831 67 788832 73 788833 79788855 83 788856 85 788889 70 788890 47 789235 36 789236 80 789237 61789239 56 789240 44 789242 81 789243 77 789244 66 789245 59 789246 46

Example 16: Effect of Modified Oligonucleotides on Human SNCA in HumanNeurons by Free Uptake, Multiple Dose

Selected modified oligonucleotides complementary to human SNCA weretested for their effects on SCNA mRNA levels in human neurons in vitroby free uptake Human IPS-cell derived neurons were plated at a densityof 35,000 cells per well and incubated with 247.00 nM, 740.70 nM, 2.22μM, 6.66 μM, or 20.00 μM oligonucleotide. After a treatment period of 5days total RNA was isolated from the cells and SNCA mRNA levels weremeasured by quantitative real-time PCR Human SNCA primer probe setRTS2621 (described hereinabove in Example 1) was used to measure mRNAlevels. SNCA mRNA levels were adjusted according to total RNA content,as measured by RIBOGREEN®. Results are presented in the tables below aspercent reduction of the amount of SNCA mRNA relative to untreatedcontrol cells. A value of 0% reduction indicates that the compound hadno effect or increased mRNA concentrations in the cell. As shown below,modified oligonucleotides complementary to human SNCA reduced the amountof human SNCA mRNA.

TABLE 76 Dose-dependent percent reduction of human SNCA mRNA in humanneurons by free uptake Compound SNCA inhibition (% reduction) Number0.247 μM 0.741 μM 2.220 μM 6.660 μM 20.000 μM IC₅₀ (μM) 709534 0 0 0 311 >20 709535 0 0 20 35 51 16.97 709536 0 12 21 55 68 7.01 709883 19 022 15 22 >20 709967 0 0 0 0 42 >20 741082 25 25 63 87 95 1.37

Example 17: Tolerability of Modified Oligonucleotides Complementary toHuman SNCA in Mice, 700 μg Dose

Modified oligonucleotides described above were tested in mice to assessthe tolerability of the oligonucleotides. Compound No. 387985,previously disclosed in WO 2012/068405 was also tested and is comparatoroligonucleotide. Wild type C57/Bl6 mice each received a single ICV doseof 700 μs of oligonucleotide listed in the table below. Each treatmentgroup consisted of 4 mice. A group of four mice received PBS as anegative control. At 3 hours post-injection, mice were evaluatedaccording to 7 different criteria. The criteria are (1) the mouse wasbright, alert, and responsive; (2) the mouse was standing or hunchedwithout stimuli; (3) the mouse showed any movement without stimuli; (4)the mouse demonstrated forward movement after it was lifted; (5) themouse demonstrated any movement after it was lifted; (6) the mouseresponded to tail pinching; (7) regular breathing. For each of the 7criteria, a mouse was given a subscore of 0 if it met the criteria and 1if it did not (the functional observational battery score or FOB). Afterall 7 criteria were evaluated, the scores were summed for each mouse andaveraged within each treatment group. The results are presented in thetable below.

TABLE 77 Tolerability scores in mice at 700 μg dose Compound No. FOB 3hour PBS 0.0 762836 0.0 762838 0.5 762839 0.8 762840 0.0 762880 5.8762899 1.8 762900 1.8 762901 0.0 762932 4.5 762952 0.0 762953 2.0 7630040.0 763052 6.5 763102 3.5 763391 0.0 763392 3.0 763393 4.0 763394 0.8763811 7.0 763812 5.5 763813 3.8 763814 3.5 763815 6.8 763817 2.0 7638183.8

TABLE 78 Tolerability scores in mice at 700 μg dose Compound No. FOB 3hour 709897 0.0 709940 1.8 740416 1.3 741073 2.5 741168 5.8 741205 0.0741229 1.0 741301 2.5 741330 3.3 762898 4.5 762949 3.8 762951 1.8 7629552.3 763035 0.8 763040 2.8 763050 1.5 763079 6.5 763084 2.0 763085 0.8763087 2.8 763088 4.5 763150 7.0 763151 6.8 763188 6.3 763196 2.0 7632163.0 763225 6.8 763281 7.0 763299 5.0 763312 2.3 763359 7.0 763384 2.3763481 5.3 763485 0.0 763650 6.8 762954 6.3

TABLE 79 Tolerability scores in mice at 700 μg dose Compound No. FOB 3hour PBS 0.0 709548 0.0 709632 6.3 740439 3.5 741018 6.8 741038 0.0762895 3.0 762896 5.0 762897 6.0 762926 3.8 762946 2.0 762947 2.8 7629481.3 762950 1.3 762956 6.0 763033 0.0

TABLE 80 Tolerability scores in mice at 700 μg dose Compound No. FOB 3hour PBS 0.0 788813 2.8 788814 0.8 788815 0.0 788816 0.0 788817 0.0788818 0.0 788819 4.8 788820 2.3 789229 0.5 789230 0.0 789232 0.0 7892330.0 789234 4.3 789235 4.3 789236 0.0 789237 0.0 789238 0.0 789239 0.0789240 6.0 789241 5.0 789242 0.3 789243 0.3 789244 0.0 789245 0.0

TABLE 81 Tolerability scores in mice at 700 μg dose Compound No. FOB 3hr PBS 0.0 762837 0.0 762901 0.0 762952 0.3 763002 0.3 763032 5.0 7630851.3 763364 1.3 763391 2.3 788833 0.0 789239 0.0 789242 0.0 789243 0.0

TABLE 82 Tolerability scores in mice at 700 μg dose PBS 0.0 387985 6.0827592 0.0 827599 0.0 827606 0.0 827607 1.0 827611 0.5 827617 5.5 8276300.0 827649 3.8 827653 3.3 827691 1.5 827695 0.0 827701 1.0 827714 0.0

Example 18: Tolerability of Modified Oligonucleotides Complementary toHuman SNCA in Rats, 3 mg Dose

Modified oligonucleotides described above were tested in rats to assessthe tolerability of the oligonucleotides. Compound No. 387985,previously disclosed in WO 2012/068405 was also tested and is comparatoroligonucleotide. Sprague Dawley rats each received a single intrathecal(IT) dose of 3 mg of oligonucleotide listed in the table below. Eachtreatment group consisted of 4 rats. A group of four rats received PBSas a negative control. At 3 hours post-injection, movement in 7different parts of the body were evaluated for each rat. The 7 bodyparts are (1) the rat's tail; (2) the rat's posterior posture; (3) therat's hind limbs; (4) the rat's hind paws; (5) the rat's forepaws; (6)the rat's anterior posture; (7) the rat's head. For each of the 7different body parts, each rat was given a sub-score of 0 if the bodypart was moving or 1 if the body part was paralyzed. After each of the 7body parts were evaluated, the sub-scores were summed for each rat andthen averaged for each group. For example, if a rat's tail, head, andall other evaluated body parts were moving 3 hours after the 3 mg ITdose, it would get a summed score of 0. If another rat was not movingits tail 3 hours after the 3 mg IT dose but all other evaluated bodyparts were moving, it would receive a score of 1. Results are presentedas the average score for each treatment group.

TABLE 83 Tolerability scores in rats at 3 mg dose Compound No. FOB 3 hrPBS 0.0 762948 2.3 762949 2.0 762951 0.8 763001 1.3 763002 2.5 7630032.5 763040 1.0 763050 3.0 763084 4.0 763085 2.0 763087 2.8 763196 3.0763233 3.3 763391 0.0

TABLE 87 Tolerability scores in rats at 3 mg dose Compound No. FOB 3 hrPBS 0.3 762946 1.5 762969 0.3 763394 3.5 763813 1.8 763817 4.5 7638183.0 789235 3.3 789236 0.8  789237* 2.0 789239 0.0 789242 1.0 789243 1.8789244 0.8 789245 0.5 *789237 group had 5 rats

TABLE 85 Tolerability scores in rats at 3 mg dose Compound No. FOB 3 hrPBS 0.0 762932 5.0 762962 6.0 763485 3.0 789231 2.5 806693 4.5 8066943.0 806695 5.0 806697 1.3 806698 4.3 806700 5.3 806701 4.0 806714 3.5806715 2.3 806716 3.5

TABLE 86 Tolerability scores in rats at 3 mg dose Compound No. FOB 3 hrPBS 0.0 806708 2.8 806709 2.0 806710 3.3 806711 3.8 806712 3.5 8067132.8

TABLE 87 Tolerability scores in rats at 3 mg dose Compound No. FOB 3 hrPBS 0.0 762836 2.3 762837 1.0 762838 0.5 762839 0.0 762840 0.0 7628953.5 762896 2.5 762899 2.8 763032 4.8 763364 3.3 788833 1.0 788890 3.0763207 2.8  806716* 3.0 *806716 group only contained 2 mice

TABLE 88 Tolerability scores in rats at 3 mg dose Compound No. FOB 3 hrPBS 0.0 762837 1.0 762901 2.8 762952 4.0 763002 3.3 763032 4.5 7630854.0 763364 4.0 763391 1.3 788833 0.8 789239 0.5 789242 3.0 789243 1.5

TABLE 89 Tolerability scores in rats at 3 mg dose Compound No. FOB 3 hrPBS 0.25 387985 3.8 827592 3.5 827599 2.0 827606 4.0 827607 1.0 8276112.5 827617 3.0 827630 1.0 827649 5.5 827653 2.4 827691 3.3 827695 1.8827701 2.5 827714 0.3

Example 19: Potency of Modified Oligonucleotides Complementary to HumanSNCA in Transgenic Mice

Modified oligonucleotides described above were tested in the SNCA PACtransgenic mouse model which uses bacterial P1 artificial chromosome(PAC) containing the entire wild-type human SNCA gene.

Treatment

The SNCA PAC mice were divided into groups of 4-8 mice each. Two groupswere tested with each compound. Groups were given a single ICV bolus ofoligonucleotide at a dose of 10, 30, 100, 300, or 700 mg and sacrificedtwo weeks later. The PBS-injected group served as the control group towhich oligonucleotide-treated groups were compared.

RNA Analysis

After two weeks, mice were sacrificed and RNA was extracted fromcortical brain tissue for real-time PCR analysis of measurement of mRNAexpression of SNCA using primer probe set hSNCA LTS00672 (forwardsequence TGGCAGAAGCAGCAGGAAA, designated herein as SEQ ID NO: 14;reverse sequence TCCTTGGTTTTGGAGCCTACA, designated herein as SEQ ID NO:15; probe sequence 5′-FAM-CAAAAGAGGGTGTTCTC-3′MGB, designated herein asSEQ ID NO: 16.). Results are presented as percent change of mRNA,relative to PBS control, normalized with cyclophilin A.

As shown in the table below, treatment with modified oligonucleotidesresulted in significant reduction of SNCA mRNA in comparison to the PBScontrol. Results are a combination of two individual studies Animalswere removed from analysis using ROUT at 1% to remove outliers. 763085had 3 animals removed with ROUT analysis and 1 animal did not survivesurgery. 763364 had 2 animals removed with ROUT analysis. 763391 had oneanimal removed with a value of 253% of control and 3 animals did notsurvive surgery. 789243 had 1 animal removed with ROUT analysis. 827599had 4 animals removed with ROUT analysis.

TABLE 90 Dose-dependent percent reduction of human SNCA mRNAintransgenic mice Cortex Compound Dose % ED50 No. (μg) Reduction (μg) PBS— — 763085 10 0 35 30 47 100 72 300 96 700 94 763364 10 35 21 30 55 10087 300 90 700 97 763391 10 17 19 30 67 100 68 300 93 700 94 789243 10 3561 30 20 100 60 300 85 700 92 827599 10 1 56 30 25 100 81 300 87 700 95

Example 20: Potency of Modified Oligonucleotides Complementary to HumanSNCA in Transgenic Mice

Modified oligonucleotides described above were tested in the SNCA PACtransgenic mouse model which uses bacterial P1 artificial chromosome(PAC) containing the entire wild-type human SNCA gene.

Treatment

The SNCA PAC mice were divided into groups of 10 mice each. Two groupswere tested with each compound. Groups were given a single ICV bolus ofoligonucleotide at a dose of 10, 30, 100, 300, or 700 mg and sacrificedtwo weeks later. The PBS-injected group served as the control group towhich oligonucleotide-treated groups were compared.

RNA Analysis

After two weeks, mice were sacrificed and RNA was extracted fromcortical brain tissue for real-time PCR analysis of measurement of mRNAexpression of SNCA using primer probe set hSNCA LTS00672 (forwardsequence TGGCAGAAGCAGCAGGAAA, designated herein as SEQ ID NO: 14;reverse sequence TCCTTGGTTTTGGAGCCTACA, designated herein as SEQ ID NO:15; probe sequence 5′-FAM-CAAAAGAGGGTGTTCTC-3′MGB, designated herein asSEQ ID NO: 16.). Results are presented as percent change of mRNA,relative to PBS control, normalized with cyclophilin A.

As shown in the table below, treatment with modified oligonucleotidesresulted in significant reduction of SNCA mRNA in comparison to the PBScontrol Animals were removed from analysis using ROUT at 1% to removeoutliers. The values in the table below are the average of 10 animalsfor all groups except the 700 μg dose, which is the average of 7 animals

TABLE 91 Dose-dependent percent reduction of human SNCA mRNAintransgenic mice Compound Dose modified oligonucleotide ED50 No. PBS 10μg 30 μg 100 μg 300 μg 700 μg (μg) 763391 0 36 51 92 94 97 21

Example 21: Potency of Modified Oligonucleotides Targeting Human SNCA inNon-Human Primates, 2 Week Study

Modified oligonucleotides described above were further evaluated forpotency in non-human primates (NHP).

Treatment

Female cynomolgus monkeys were divided into groups of 4 NHP each. Groupsreceived a single IT bolus of 35 mg of modified oligonucleotide 789243,763391, 763364, 763085, or 827599. One group of NHP received a dose ofartificial cerebrospinal fluid (aCSF). The aCSF-injected group served asthe control group to which oligonucleotide-treated groups were compared.After two weeks, NHP were sacrificed and tissues were collected foranalysis.

RNA Analysis

RNA was extracted from various neural tissues for real-time PCR analysisof mRNA expression of SNCA as in the previous example. Results arepresented as percent change of mRNA, relative to aCSF control,normalized with NHP Cyclophylin A. As shown in the table below,treatment with modified oligonucleotides resulted in reduction of SNCAmRNA in comparison to the PBS control with some of the treatment groups.The lumbar cord is an average of 3 NHP for 763391 because one lumbarsample was only able to obtain cauda aquina, and thus not from thelumbar region.

TABLE 92 Reduction of human SNCA mRNA in Non-Human Primates % ReductionLumbar Compound Spinal Frontal Thoracic Temporal No Cord Cortex cordcortex aCSF 0 0 0 0 763085* 22 9 9 5 763364* 0 10 0 10 763391  85 50 5945 789243  38 34 26 3 827599  51 29 32 28 *These two oligos each containsingle mismatch to cynomolgus SNCA

Example 22: Potency of Modified Oligonucleotides Targeting Human SNCA inNon-Human Primates, 13 Week Study

Modified oligonucleotides described above were further evaluated forpotency and tolerability in non-human primates (NHP).

Treatment

Female cynomolgus monkeys were divided into groups of 4 NHP each. Groupsreceived an IT bolus dose of 35 mg of Compound 763391 or Compound 827599on day one, on day 14, and then monthly for a total of 5 doses. Onegroup of NHP received doses of aCSF rather than oligonucleotide. TheaCSF-injected group served as the control group to whicholigonucleotide-treated groups were compared. A week after the finaldose, NHP were sacrificed and tissues were collected for analysis.

RNA Analysis

RNA was extracted from various neural tissues for real-time PCR analysisof mRNA expression of SNCA as in examples above. Results are presentedas percent change of mRNA, relative to aCSF control, normalized withmonkey Cyclophylin A.

As shown in the table below, treatment with modified oligonucleotidesresulted in reduction of SNCA mRNA in comparison to the PBS control.

TABLE 93 Reduction of human SNCA mRNA in Non-Human Primates % ReductionMidbrain Cervical Thoracic Lumbar (level of Comp Spinal Spinal SpinalMotor Frontal Substantia No Cord Cord Cord Cortex Cortex CaudateAmygdala Pons nigra) Putamen aCSF 0 0 0 0 0 0 0 0 0 0 763391 96 96 97 9898 57 98 77 83 9 827599 68 80 95 61 72 27 70 41 31 0

TABLE 94 Reduction of human SNCA mRNA in Cynomolgus Monkeys % ReductionMedulla Comp. Cerebellar Corpus Ent Hippo Insular (rostral and CaudalCentml No Ped-uncle Collo-sum DCN Cortex campus Hypo Cortex dorsal)Medulla Gray aCSF 0 0 0 0 0 0 0 0 0 0 763391 90 91 47 97 90 78 97 74 9068 827599 61 67 37 71 25 25 68 38 54 9

TABLE 95 Reduction of human SNCA mRNA in Cynomolgus Monkeys % ReductionDorsal Rostral Comp VPM/ Occipital Cerebral Temporal Medial Globus VA/Ventral Superior No VPL Pulvinar cortex cortex Cortex Thalamus PallidusVL Medulla Colliculi aCSF 0 0 0 0 0 0 0 0 0 0 763391 63 96 96 58 97 1256 45 84 68 827599 26 59 58 0 72 0 0 13 52 9

Example 23: Treatment of SNCA Pathology in Pre-Formed Fibril (PFF) Modelin Wild-Type Mouse, Prophylactic Treatment Experimental Model

The PFF (pre-formed fibril) model in mouse is an experimental model thathas been used to investigate treatments for Parkinson's disease, asdescribed in Luk, et. al., Science. 2012 Nov. 16; 338(6109):949-53. Asingle intrastriatal injection of pre-formed SNCA fibrils generates Lewybody pathology characteristic of Parkinson's disease.

Modified Oligonucleotide

Compound No: 678363 is a 4-8-5 MOE and cEt gapmer that is 100%complementary to mouse SNCA, having a sequence of (from 5′ to 3′)TTTAATTACTTCCACCA (incorporated herein as SEQ ID NO:23), having a sugarmotif of (from 5′ to 3′): eeekddddddddkeeee; wherein ‘d’ represents a2′-deoxyribose sugar; ‘e’ represents a 2′-MOE modified sugar; and ‘k’represents a cEt modified sugar; and an internucleoside linkage motif of(from 5′ to 3′) soosssssssssooss; wherein ‘o’ represents aphosphodiester internucleoside linkage and ‘s’ represents aphosphorothioate internucleoside linkage.

Experimental Protocol

Three groups of twelve mice wild-type B6C3F1 mice were treated accordingto the table below. 700 μs modified oligonucleotide or PBS wasadministered via ICV (intracerebroventricial) injection on day 0, andpre-formed fibrils were administered into the striatum on day 14. On day56, a wirehang test was performed to measure motor function, and micewere sacrificed for mRNA and histological analysis. P-α-Syn aggregatesin the substantia nigra were stained and quantified. Mouse SNCA mRNA wasmeasured with RT-PCR as described above, using mouse primer probe setRTS2956 (forward sequence GTCATTGCACCCAATCTCCTAAG, designated herein asSEQ ID NO: 17; reverse sequence GACTGGGCACATTGGAACTGA, designated hereinas SEQ ID NO: 18; probe sequence CGGCTGCTCTTCCATGGCGTACAA, designatedherein as SEQ ID: 19). SNCA mRNA levels were normalized to cyclophilin Aand are presented as % of mRNA level in PBS-treated mice. As shown inthe table below, modified oligonucleotide treated mice had reduced SNCAmRNA, fewer aggregates in substantia nigra and improved performance onthe wirehang test as compared to PBS treated mice.

TABLE 96 Pre-formed fibril (PFF) model in wild-type mouse, prophylactictreatment Treatment SNCA mRNA Avg. # aggregates in Wirehang Treatmentgroup Day 0 Day 14 Midbrain striatum substantia nigra (seconds) Naïve(Naïve) PBS none 90 98 0 192 ± 82 PBS + PFF (PBS) PBS PFF 100 100 42  94± 69 678363 + PFF 678363 PFF 51 47 0.64 154 ± 64 (Treatment)

Example 24: Treatment of SNCA Pathology in Pre-Formed Fibril (PFF) Modelin Mouse, Post-Symptomatic Treatment Experimental Protocol

Three groups of twelve wild-type B6C₃F1 mice were treated according tothe table below. Pre-formed fibrils were administered into the striatumon day 0 and 700 μg modified oligonucleotide or PBS was administered viaICV (intracerebroventricial) injection on day 14. On day 56, a wirehangtest was performed to measure motor function, and mice were sacrificedfor mRNA and histological analysis. Phosphorylated-α-Syn aggregates inthe substantia nigra were stained and quantified. Mouse SNCA mRNA wasmeasured as in the previous example and normalized to PBS treated mice.As shown in the table below, modified oligonucleotide treated mice hadreduced SNCA mRNA and fewer aggregates in substantia nigra and improvedperformance on the wirehang test as compared to PBS treated mice.

TABLE 97 Pre-formed fibril (PFF) model in wild-type mouse,post-symptomatic treatment Treatment SNCA mRNA Avg. # p-αSyn Treatmentgroup Day 0 Day 14 Midbrain striatum aggregates in SN Wirehang (s) Naïve(Naïve) PBS none 102 85 0 226 ± 88 PFF + PBS (PBS) PFF PBS 100 100 49.4 58 ± 63 PFF + 678363 PFF 678363 32 32 1.9 132 ± 77 (Treatment)

Example 25: Treatment of SNCA Pathology in Pre-Formed Fibril (PFF) Modelin Mouse, Long-Term Prophylactic Treatment Experimental Protocol

Three groups of twelve mice wild-type B6C3F1 mice were treated accordingto the table below. 700 μg modified oligonucleotide (control ortreatment) or PBS was administered via ICV (intracerebroventricial)injection on day 0, pre-formed fibrils were administered into thestriatum on day 14, and an additional 700 μg modified oligonucleotide orPBS was administered via ICV on day 90.

Control groups included a PBS-treated group and Compound No.676630-treated group. Compound No: 676630 is a 5-10-5 MOE gapmer that isnot complementary to mouse SNCA, having a sequence of (from 5′ to 3′)CCTATAGGACTATCCAGGAA (incorporated herein as SEQ ID NO: 2795) and havingan internucleoside linkage motif of (from 5′ to 3′) sooosssssssssssoos,wherein ‘o’ represents a phosphodiester internucleoside linkage and ‘s’represents a phosphorothioate internucleoside linkage.

On day 180, mice were sacrificed for mRNA and histological analysis.Phosphorylated-α-Syn aggregates and neurtic pathology in the substantianigra were stained and quantified for 6 mice in each group.Additionally, the number of TH (tyrosine hydroxylase)+cells in thesubstantia nigra pars compacta (SNpc), which is measure of death ofdopaminergic neurons, were quantified for 6 mice in each group. Resultsare presented relative to the PBS-treated group. As shown in the tablebelow, Compound No. 677363-treated mice had reduced SNCA mRNA, feweraggregates in substantia nigra, and reduced neuritic pathology insubstantia nigra as compared to PBS and 676630-treated mice.

TABLE 98 Pre-formed fibril (PFF) model in wild-type mouse, long-termprophylactic treatment Treatment SNCA mRNA Avg. # p-αSyn Avg. # neuriticTH cells Treatment Day Day Mid aggregates pathology (% PBS) group Day 014 90 brain striatum in SN in SN in SNpc PFF + PBS PBS PFF PBS 100 100271 1002 100 676630 676630 PFF 676630 95 107  160*   580* 99 (Control)678363 678363 PFF 678363 59 70    0.7  51 139 (Treatment) *numberrepresents the average from 4 mice.

Example 26: Tolerability of Modified Oligonucleotides Complementary toHuman SNCA in Mice, 700 μg Dose

Modified oligonucleotides described above were tested against CompoundNos. 1233344 and 1233345 (described herein below) to assess thetolerability of the oligonucleotides.

Compound No. 1233344 is a 15-mer gapmer, that is complementary to SNCA(wherein the 5′-most nucleoside to which the gapmer targets SEQ ID NO: 1is at position 370), having a sequence of (from 5′ to 3′)CTACATAGAGAACAC (incorporated herein as SEQ ID No.:2796), wherein eachof the nucleosides 1-3, nucleoside 13 and nucleoside IA (from 5′ to 3′)comprise an LNA sugar modification, and each of the nucleosides 4-12 andnucleoside 15 are deoxynucleosides, wherein the internucleoside linkagesbetween the nucleosides are phosphorothioate internucleoside linkages.Compound No. 1233344 is characterized by the following chemicalnotation:C_(luas)T_(lnas)A_(lnas)C_(ds)A_(dk)T_(ds)A_(ch)G_(ds)A_(ds)G_(ds)A_(ds)A_(ck)C_(luas)A_(luas)C_(d)wherein,

A=an adenine, nucleobase,

C=a cytosine nucleobase,

G=a guanine nucleobase.

T=a thymine nucleobase,

d=a 2′-deoxyribose sugar,

s===a phosphorothioate internucleoside linkage, and

lna===an LNA modified sugar.

Compound No. 1233345 is a 15-mer gapmer, that is complementary to SNCA(wherein the 5′-most nucleoside to which the gapmer targets SEQ ID NO: 1is at position 372), having a sequence of (from 5′ to 3′)GCCTACATAGAGAAC (incorporated herein as SEQ ID No.:2797), wherein eachof the nucleosides 1-3, nucleoside 13 and nucleoside IA (from 5′ to 3′)comprise an LNA sugar modification, and each of the nucleosides 4-12 andnucleoside 15 are deoxy nucleosides, wherein the internucleosidelinkages between the nucleosides are phosphorothioate internucleosidelinkages. Compound No. 1233345 is characterized by the followingchemical notation:G_(lnas)C_(luas)C_(luas)T_(ds)A_(ds)C_(ds)A_(ds)T_(ds)A_(ds)G_(ds)A_(ds)G_(ds)A_(lnas)A_(lnas)C_(d)wherein,

A=an adenine, nucleobase,

C=a cytosine nucleobase,

G=a guanine nucleobase.

T=a thymine nucleobase,

d=a 2′-deoxyribose sugar,

s a phosphorothioate internucleoside linkage, and

lna=an LNA modified sugar.

Treatment

Wildtype C57BL/6 mice each received a single ICV dose of 700 μg ofmodified oligonucleotide listed in the table below. Each treatment groupconsisted of 4 mice. A group of four mice received PBS as a negativecontrol. At 3 hours post-injection, mice were evaluated according to 7different criteria. The criteria are (1) the mouse was bright, alert,and responsive; (2) the mouse was standing or hunched without stimuli;(3) the mouse showed any movement without stimuli; (4) the mousedemonstrated forward movement after it was lifted; (5) the mousedemonstrated any movement after it was lifted; (6) the mouse respondedto tail pinching; (7) regular breathing. For each of the 7 criteria, amouse was given a subscore of 0 if it met the criteria and 1 if it didnot (the functional observational battery score or FOB). After all the 7criteria were evaluated, the scores were summed for each mouse andaveraged within each treatment group. The results are presented in thetable below.

TABLE 99 Tolerability scores in mice at 700 μg dose Compound No. FOB 3hour PBS 0.0 763085 2.3 763364 2.8 763391 0.0 789243 1.0 827599 1.01233345 6.8 1233344 2.3

1-80. (canceled)
 81. An oligomeric compound comprising a modifiedoligonucleotide consisting of 10-30 linked nucleosides and having anucleobase sequence comprising at least 12 nucleobases of any of SEQ IDNOS: 28-2793.
 82. An oligomeric compound comprising a modifiedoligonucleotide consisting of 10-30 linked nucleosides and having anucleobase sequence complementary to at least 8, at least 9, at least10, at least 11, at least 12, at least 13, at least 14, at least 15, atleast 16, at least 17, at least 18, at least 19, or at least 20contiguous nucleobases of: (i) an equal length portion of nucleobases50915-50943 of SEQ ID NO: 2; (ii) an equal length portion of nucleobases19630-19656 of SEQ ID NO: 2; (iii) an equal length portion ofnucleobases 28451-28491 of SEQ ID NO: 2; (iv) an equal length portion ofnucleobases 48712-48760 of SEQ ID NO: 2; (v) an equal length portion ofnucleobases 23279-23315 of SEQ ID NO: 2; (vi) an equal length portion ofnucleobases 20964-21018 of SEQ ID NO: 2; (vii) an equal length portionof nucleobases 22454-22477 of SEQ ID NO: 2; (viii) an equal lengthportion of nucleobases 72294-72321 of SEQ ID NO: 2; (ix) an equal lengthportion of nucleobases 20549-20581 of SEQ ID NO: 2; or (x) an equallength portion of nucleobases 27412-27432 of SEQ ID NO:
 2. 83. Theoligomeric compound of claim 81, wherein the modified oligonucleotidecomprises at least one modified nucleobase.
 84. The oligomeric compoundof claim 83, wherein the modified nucleobase is a 5-methyl cytosine. 85.The oligomeric compound of claim 81, wherein the modifiedoligonucleotide consists of 17 or 20 linked nucleosides.
 86. Theoligomeric compound of claim 81, wherein the modified oligonucleotidecomprises at least one modified nucleoside comprising a modified sugarmoiety.
 87. The oligomeric compound of claim 86, wherein the modifiedoligonucleotide comprises at least one modified nucleoside comprising abicyclic sugar moiety.
 88. The oligomeric compound of claim 87, whereinthe modified oligonucleotide comprises at least one modified nucleosidecomprising a bicyclic sugar moiety having a 2′-4′ bridge, wherein the2′-4′ bridge is selected from —O—CH₂— and —O—CH(CH₃)—.
 89. Theoligomeric compound of claim 86, wherein the modified oligonucleotidecomprises at least one modified nucleoside comprising a non-bicyclicmodified sugar moiety.
 90. The oligomeric compound of claim 89, whereinthe modified oligonucleotide comprises at least one modified nucleosidecomprising a non-bicyclic modified sugar moiety comprising a 2′-MOEmodified sugar or a 2′-OMe modified sugar.
 91. The oligomeric compoundof claim 86, wherein the modified oligonucleotide has a sugar motifcomprising: a 5′-region consisting of 1-5 linked 5′-region nucleosides;a central region consisting of 7-12 linked central region nucleosides;and a 3′-region consisting of 1-5 linked 3′-region nucleosides; whereinthe 3′-most nucleoside of the 5′-region and the 5′-most nucleoside ofthe 3′-region comprise modified sugar moieties, and each of the centralregion nucleosides is a 2′-β-D-deoxyribosyl sugar moiety.
 92. Theoligomeric compound of claim 91, wherein the modified oligonucleotideconsists of 17 linked nucleosides; the central region of the modifiedoligonucleotide comprises at least 9 linked central region nucleosides;the modified oligonucleotide comprises at least four bicyclicnucleosides; each of the at least four bicyclic nucleosides comprises abicyclic sugar moiety having a 2′-4′ bridge, wherein the 2′-4′ bridge isselected from —O—CH₂— and —O—CH(CH₃).
 93. The oligomeric compound ofclaim 81, wherein the modified oligonucleotide comprises at least onemodified internucleoside linkage.
 94. The oligomeric compound of claim93, wherein each internucleoside linkage of the modified oligonucleotideis a modified internucleoside linkage.
 95. The oligomeric compound ofclaim 93 wherein at least one modified internucleoside linkage is aphosphorothioate internucleoside linkage.
 96. The oligomeric compound ofclaim 94, wherein each internucleoside linkage of the modifiedoligonucleotide is a phosphorothioate internucleoside linkage.
 97. Theoligomeric compound of claim 92, wherein each internucleoside linkage ofthe modified oligonucleotide is a phosphorothioate internucleosidelinkage.
 98. The oligomeric compound of claim 81, which consists of themodified oligonucleotide.
 99. The oligomeric compound of claim 81,wherein the oligomeric compound comprises a conjugate group comprising aconjugate moiety and a conjugate linker.
 100. The oligomeric compound ofclaim 81 wherein the oligomeric compound is a single-stranded oligomericcompound.
 101. An oligomeric duplex comprising an oligomeric compound ofclaim
 81. 102. A population of oligomeric compounds of claim 95, whereinall of the phosphorothioate internucleoside linkages of the modifiedoligonucleotide are stereorandom.
 103. A pharmaceutical compositioncomprising an oligomeric compound of claim 81 and a pharmaceuticallyacceptable diluent.
 104. The pharmaceutical composition of claim 103,wherein the pharmaceutically acceptable diluent is phosphate-bufferedsaline (PBS) or artificial cerebrospinal fluid.
 105. The pharmaceuticalcomposition of claim 104, wherein the pharmaceutical compositionconsists essentially of the oligomeric compound and PBS or artificialcerebrospinal fluid.
 106. A method comprising administering to an animala pharmaceutical composition of claim
 103. 107. A method of treating adisease associated with SNCA comprising administering to an individualhaving or at risk for developing a disease associated with SNCA atherapeutically effective amount of a pharmaceutical composition ofclaim 103 and thereby treating the disease associated with SNCA. 108.The method of claim 107, wherein the disease associated with SNCA is aneurodegenerative disease.
 109. The method of claim 108, wherein theneurodegenerative disease is Parkinson's disease, dementia with Lewybodies, diffuse Lewy body disease, pure autonomic failure, multiplesystem atrophy, neuronopathic Gaucher's disease, or Alzheimer's disease.110. The method of claim 109, wherein at least one symptom or hallmarkof the neurodegenerative disease is ameliorated.
 111. The method ofclaim 110, wherein the symptom or hallmark is motor dysfunction,aggregation of alpha-synuclein, neurodegeneration, cognitive decline,and/or dementia.